Immunogenicity of Novel Monovalent Oral Poliovirus Vaccine Type 2 (nOPV2) With and Without Bivalent OPV
Phase: PHASE2
Conditions: Poliomyelitis
Sponsor: Centers for Disease Control and Prevention

 
1 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
 
 
 
 
RRC APPLICATION FORM 
RESEARCH PROTOCOL  
Number: 20060  
Version No. 1.06 
Version date: 03-Dec-2020 
 
FOR OFFICE USE ONLY 
RRC Approval: 
 Yes 
 No 
Date:17.07.2020 
ERC Approval: 
 Yes 
 No 
Date: 22.08.2020 
AEEC Approval: 
 Yes 
 No 
Date:      
External IRB Approval  
 Yes 
 No      Date:      
Name of External IRB: ___________________________________________ 
Protocol Title:* (maximum 250 characters including space) Immunogenicity of novel monovalent oral poliovirus 
vaccine type 2 (nOPV2) with and without bivalent OPV 
 
Short Title: (maximum 100 characters including space)  
nOPV2 with and without bOPV 
 
Key Words:* nOPV2, bOPV, immunogenicity, Bangladesh 
Name of the Research Division Hosting the Protocol:* 
  Health Systems and Population Studies Division (HSPSD) 
  Nutrition and Clinical Services Division (NCSD) 
  Infectious Diseases Division (IDD) 
 
 
   Maternal and Child Health Division (MCHD) 
   Laboratory Sciences and Services Division (LSSD) 
   Other (specify) ____________________________ 
Has the Protocol been Derived from an Activity:*   
  No            
 Yes (please provide following information): 
Activity No.  :       
Activity Title:       
PI:       
Grant No.:        
Budget Code:       
 
Start Date:        
End Date:       
icddr,b Strategic Priority/ Initiative (SP 2015-8):* (check all that apply)  
   Reducing maternal and neonatal mortality 
   Controlling enteric and respiratory infections 
   Preventing and treating maternal and childhood malnutrition 
   Detecting and controlling emerging and re-emerging infections 
   Achieving universal health coverage 
   Examining the health consequences of climate change 
   Preventing and treating non-communicable diseases 
   Others (specify) ________________________________ 
Research Phase (4 Ds):* (check all that apply) 
   Discovery 
   Development 
 
    Delivery 
   Evaluation of Delivery 
Anticipated Impact of Research:* (check all that apply and 
please provide details below) 
   Knowledge Production 
   Capacity Building 
 
 
 
 
 
 
 
 
   Informing Policy 
   Health and Health Sector Benefits 
   Economic Benefits 
 
 

 
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Please provide details here:  
 
The results will guide Global Polio Eradication Initiative (GPEI), including WHO, in making policy for 
responding to polio outbreaks with the goal of eradication of polio. 
 
 
 
Which of the Sustainable Development Goal This Protocol Relates to?:* (check all that apply)                              
 1. End poverty in all its forms everywhere 
 2. End hunger, achieve food security and improved nutrition and promote sustainable agriculture 
 3. Ensure healthy lives and promote well-being for all at all ages 
 4. Ensure inclusive and equitable quality education and promote lifelong learning opportunities for all 
 5. Achieve gender equality and empower all women and girls 
 6. Ensure availability and sustainable management of water and sanitation for all 
 7. Ensure access to affordable, reliable, sustainable and modern energy for all 
 8. Promote sustained, inclusive and sustainable economic growth, full and productive employment and decent work for all 
 9. Build resilient infrastructure, promote inclusive and sustainable industrialization and foster innovation 
 10. Reduce inequality within and among countries 
 11. Make cities and human settlements inclusive, safe, resilient and sustainable 
 12. Ensure sustainable consumption and production patterns 
 13. Take urgent action to combat climate change and its impacts 
 14. Conserve and sustainably use the oceans, seas and marine resources for sustainable development 
 15. Protect, restore and promote sustainable use of terrestrial ecosystems, sustainably manage forests, combat 
desertification, and halt and reverse land degradation and halt biodiversity loss 
 16. Promote peaceful and inclusive societies for sustainable development, provide access to justice for all and build 
effective, accountable and inclusive institutions at all levels 
 17. Strengthen the means of implementation and revitalize the global partnership for sustainable development 
Does this Protocol Use the Gender Framework:*                         
(Please visit: http://www.icddrb.net.bd/jahia/Jahia/pid/684  for 
Gender Alanysis Tool with instructions) 
  Yes (please complete Gender Analysis Tool) 
  No 
If ‘no’ is the response, its reason(s) in brief: Nothing known about gender issues in polio vaccination 
Will this Research Specifically Benefit the Disadvantaged (economically, socially and/or 
otherwise): 
 Yes 
   No      
Does this Protocol use Behaviour Change Communication:     
  Yes 
  No     
Principal Investigator (Should be icddr,b staff):*    Sex   
  Female    
   Male 
K. Zaman, Senior Scientist and Epidemiologist  
kzaman@icddrb.org Exg: 3806 
Cell phone: 01713047100, 
Do you have ethics certification?  
 No   
  Yes (please attach in your CV below) 
Do you have RBM training certification?   
 No 
 Yes (please attach the 
certificate with CV below) 
 Primary Scientific Division of the PI 
 
IDD 
Co-Principal Investigator(s) Internal:  Sex    
  Female    
  Male 
Md. Yunus, myunus@icddrb.org 
 
Signature or written consent of Co-PI: __________________ 
(electronic signature or email or any sort of written consent) 
[if more than one, please copy and paste this row for additional Co-PIs]  
 
Do you have ethics certification?   
 No    
  Yes (please attach in your CV 
below) 
Do you have RBM training certification?   
 No 
  Yes (please attach the 
certificate with CV below) 
Primary Scientific Division/ 
Programme  of the Co-PI 
 
E-mail approval Page 107 
 
HSPSD 
___________________________ 
Approval of the Respective Senior 
Director/ Programme Head 
 
 
 
 
(Signature) 

 
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Co-Investigator(s) - Internal:  Sex    
  Female    
  Male 
Dr. Asma Binte Aziz 
(Position, phone no, extension no, cell, and email address ):  
Research Investigator, asma.aziz@icddrb.org 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) 
[if more than one, please copy and paste this row for additional Co-Is 
 
Do you have ethics certification?      
 No    
 Yes (please attach in your CV 
below) 
Do you have RBM training certification?   
 No 
 Yes (please attach the certificate 
with CV below) 
Primary Scientific Division of the 
Co-I  
 IDD 
 
E-mail approval Page 107 
 
 
___________________________ 
Approval of the Respective Senior 
Director/ Programme Head 
 
 
 
(Signature) 
Co-Investigator(s) - Internal:  Sex    
  Female    
  Male 
Dr. Masuma Hoque 
(Position, phone no, extension no, cell, and email address ):  
 Senior Research Investigator, masuma.hoque@icddrb.org 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) 
[if more than one, please copy and paste this row for additional Co-Is 
 
Do you have ethics certification?      
 No    
 Yes (please attach in your CV 
below) 
Do you have RBM training certification?   
 No 
 Yes (please attach the certificate 
with CV below) 
Primary Scientific Division of the 
Co-I   
               IDD 
 
E-mail approval Page 108 
 
 
___________________________ 
Approval of the Respective Senior 
Director/ Programme Head 
 
 
 
(Signature) 
Co-Principal Investigator(s) - External: Sex    
 Female     
   Male 
Amanda L. Wilkinson  
Global Immunization Division 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
lxq6@cdc.gov  
 
 
Signature or written consent of Co-PI: __________________ 
(electronic signature or email or any sort of written consent)  E-mail approval Page 122 
 
Co-Investigator(s) - External:  Sex    
  Female    
  Male 
Abhijeet Anand 
Global Immunization Division 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
aanand@cdc.gov  
office: +1-404-639-1970 
Blackberry: +1-404-797-9074 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 122 
 
 

 
4 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Co-Investigator(s) - External:  Sex    
  Female    
  Male 
Cindi Snider 
Global Immunization Division 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
csnider@cdc.gov  
Office: +1-404-718-6328 
Blackberry: +1-404-431-4626 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 123 
 
 
Co-Investigator(s) – External:  Sex  
  Female     
  Male 
Concepcion F. Estivariz 
Global Immunization Division 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
cge3@cdc.gov 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 124 
 
 
Co-Investigator(s) – External:  Sex  
  Female     
  Male 
Qian An  
Global Immunization Division 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
fei8@cdc.gov   
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 125 
 
Co-Investigator(s) – External:  Sex  
  Female     
  Male 
Mark Pallansch 
National Center for Immunization and Respiratory DiseasesUS Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
map1@cdc.gov  
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 125 
 
Co-Investigator(s) – External:  Sex  
  Female     
  Male 
Steven Wassilak 
Global Immunization Division 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
sgw1@cdc.gov 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 126 
 
 

 
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Co-Investigator(s) – External:  Sex  
  Female     
  Male 
Steve Oberste 
Division of Viral Diseases 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
mbo2@cdc.gov 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 127 
 
Co-Investigator(s) - External:  Sex    
  Female    
  Male 
Jacquelyn S. Lickness 
Global Immunization Division 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
wxx3@cdc.gov 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 127 
 
 
Co-Investigator(s) - External:  Sex    
  Female    
  Male  
Jaymin Patel 
Global Immunization Division 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
isr0@cdc.gov 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent)  E-mail approval Page 128 
 
Co-Investigator(s) - External:  Sex    
  Female    
  Male  
Jennifer Anstadt  
Division of Viral Diseases 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
yrq1@cdc.gov  
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent) E-mail approval Page 129 
  
Co-Investigator(s) - External:  Sex    
  Female    
  Male  
Cara Burns 
Division of Viral Diseases 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, USA 
zqd1@cdc.gov 
 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent)  E-mail approval Page 130 
 

 
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Technical Advisor(s)  HOUSEHOLD TRANSMISSION ADD-ON COMPONENT ONLY 
 
Ananda S. Bandyopadhyay 
Bill and Melinda Gates Foundation 
500 5th Avenue N 
Seattle, WA, USA 
Ananda.Bandyopadhyay@gatesfoundation.org     
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent)  E-mail approval Page 130 
 
 
Jillian Gauld 
Institute for Disease Modeling 
139th Avenue SE 
Bellvue, WA, USA 
jgauld@idmod.org 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent)  E-mail approval Page 130 
 
 
Mike Famulare  
Institute for Disease Modeling 
139th Avenue SE 
Bellvue, WA, USA 
mfamulare@idmod.org 
Signature or written consent of Co-I: __________________ 
(electronic signature or email or any sort of written consent)  E-mail approval Page 130 
 
 

 
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Collaborating Institute(s): Please provide full official address 
 
Institution # 1 
 
Country 
USA 
Contact person 
Abhijeet Anand 
Department 
(including Division, Centre, Unit) 
Global Immunization Division 
Institution 
(with official address) 
US Centers for Disease Control and Prevention 
1600 Clifton Road NE 
Atlanta, GA, 30329 
Directorate 
(in case of GoB i.e. DGHS) 
      
Ministry (in case of GoB) 
      
 
Institution # 2 
 
Country 
Bangladesh 
Contact person 
Dr. Md. Shamsul Haque 
Department 
(including Division, Centre, Unit) 
DGHS 
Institution 
(with official address) 
 
Line Director, MNC & AH 
Directorate 
(in case of GoB i.e. DGHS) 
      
Ministry (in case of GoB) 
      
 
Note: If less than or more than three collaborating institutions, please delete or insert blocks as needed. 
 

 
8 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Contribution by the Members of the Scientific Team: 
 
 
Members’ Name 
Contribution 
Research 
idea/ 
concept 
Study 
design 
Protocol 
writing 
Respond 
to external 
reviewers’ 
comments 
Defending 
at IRB 
Developing 
data 
collection 
Tool(s) 
Data 
Collection 
Data analysis/ 
interpretation 
of results 
Manuscript  
writing 
K. Zaman 
 
 
 
 
 
 
 
 
 
Md. Yunus 
 
 
 
 
 
 
 
 
 
Amanda Wilkinson 
 
 
 
 
 
 
 
 
 
Abhijeet Anand 
 
 
 
 
 
 
 
 
 
Cindi Snider 
 
 
 
 
 
 
 
 
 
Concepcion 
Estivariz 
 
 
 
 
 
 
 
 
 
Qian An  
 
 
 
 
 
 
 
 
 
Steven Wassilak 
 
 
 
 
 
 
 
 
 
Mark Pallansch 
 
 
 
 
 
 
 
 
 
Steve Oberste 
 
 
 
 
 
 
 
 
 
Jacquelyn S. 
Lickness 
 
 
 
 
 
 
 
 
 
Asma Aziz 
 
 
 
 
 
 
 
 
 
Masuma Hoque 
 
 
 
 
 
 
 
 
 
Jaymin Patel 
 
 
 
 
 
 
 
 
 
Jennifer Anstadt 
 
 
 
 
 
 
 
 
 
Cara Burns 
 
 
 
 
 
 
 
 
 
Ananda S. 
Bandyopadhyay 
 
 
 
 
 
 
 
 
 
Jillian Gauld 
 
 
 
 
 
 
 
 
 
Mike Famulare  
 
 
 
 
 
 
 
 
 
Study Population: Sex, Age, Special Group and Ethnicity 
 
Research Subject: 
   Human             
   Animal  
 
   Microorganism 
   Other (specify): _____________ 
 
Sex: 
  Male             
  Female  
   Transgender 
 
Age: 
   0 – 4 years 
   5 – 10 years 
   11 – 17 years 
   18 – 64 years 
   65 + 
Special Group: 
   Pregnant Women 
   Fetuses 
   Prisoners 
   Destitutes  
   Service Providers 
   Cognitively Impaired 
   CSW 
   Expatriates 
   Immigrants 
   Refugee 
  Others (specify): ________________ 
 
Ethnicity: 
  No ethnic selection (Bangladeshi) 
  Bangalee 
  Tribal group 
   Other (specify): ________________ 
 
NOTE: It is icddr.b’s policy to include men, women, children and transgender in its research projects involving participation 
of humans, unless there is strong justification(s) for their exclusion.  

 
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Consent Process: (Check all that apply) 
 
  Written 
  Oral 
  Audio 
  Video 
  None 
Language: 
   Bangla 
   English 
   Other (specify: _______________ 
 
Project/Study Site: (Check all that apply) 
  Chakaria  
  Bandarban 
  Dhaka Hospital  
  Kamalapur Field Site/HDSS  
  Mirpur (Dhaka) 
  Matlab DSS Area 
  Matlab non-DSS Area 
  Matlab Hospital 
  Mirzapur 
 
 
   Bianibazar (Sylhet) 
   Kanaighat (Sylhet) 
   Jakigonj (Sylhet) 
   Other community in Dhaka 
        Name: CTU, Mohakhali_________________          
   Other sites in Bangladesh 
        Name: _________________________________ 
   Multi-national Study 
        Name of the country______________________   
Project/Study Type: (Check all that apply) 
 
  Case Control Study 
  Clinical Trial (Hospital/Clinic/Field)* 
  Community-based Trial/Intervention 
  Cross Sectional Survey 
  Family Follow-up Study 
  Longitudinal Study (cohort or follow-up) 
  Meta-analysis 
  Programme Evaluation 
 
   Programme (Umbrella Project) 
   Prophylactic Trial 
   Record Review 
   Secondary Data Analysis 
        Protocol No. of  Data Source: _____________ 
   Surveillance/Monitoring 
   Systematic Review 
   Other (specify): _________________________ 
 
 *Note: International Committee of Medical Journal Editors (ICMJE) defines Clinical Trial as “Any research project that 
prospectively assigns human participants to intervention and comparison groups to study the cause-and-effect 
relationship between a medical intervention and a health outcome”. 
PI of the RRC- and ERC-approved Clinical Trials should provide necessary information to IRB Secretariat (Research 
Administration) for registration and uploading into relevant websites (usually at the https://register.clinicaltrials.gov/). 
They should also provide relevant information to the IRB Secretariat in the event of amendment/modification after 
their approval by RRC and ERC.           
Biological Specimen: 
 
a)  Will the biological specimen be stored for future use?          
 Yes            
 No            
 Not applicable 
b)  If the response is ‘yes’, how long the specimens will be 
preserved?     
_____5_____  years 
c) What types of tests will be carried out with the preserved 
specimens? 
Studies involving polio and/or other vaccine 
preventable diseases may request preserved serum 
and/or stool specimens. No DNA or genetic testing 
of blood specimens will be permitted. Proposals to 
test stored specimens will need to be reviewed and 
approved by icddr,b’s research and ethical review 
committees as well as the institution requesting 
preserved specimens. 
d) Will the consent be obtained from the study participants for use 
of the preserved specimen for other initiative(s) unrelated to this 
study, without their re-consent? 
 Yes            
 No            
 Not applicable 
e) Will the specimens be shipped to other country/ countries? 
If yes, name of institution(s) and country/countries.   
 Yes            
 No            
 Not applicable 
Name  Centers for Disease Control and Prevention, 
Atlanta, USA___________________________ 

 
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f) 
If shipped to another country, will the surplus/unused specimen 
be returned to icddr,b?   
If the response is ‘no’, then the surplus/unused specimen must 
be destroyed. 
 Yes            
 No            
 Not applicable 
g) Who will be the custodian of the specimen at icddr,b?  
Dr. K. Zaman 
h) Who will be the custodian of the specimen when shipped 
outside Bangladesh?    
Dr. Bernardo A. Mainou, CDC 
i) 
Who will be the owner(s) of the specimens?        
icddr,b 
j) 
Has a MoU been signed with regards to collection, storage, use 
and ownership of specimen?                                    
If the response is ‘yes’, please attach a copy of the MoU.   
       If the response is ‘no’, appropriate justification should be 
provided for not signing a MoU. 
 Yes            
 No            
 Not applicable 
 
The MOU for icddr,b will be obtained prior to start of 
the study.  
Proposed Sample Size: 
 
Sub-group (Name of subgroup e.g. Men, Women) and Number 
 
Name 
Number 
Name 
Number 
(1) A: nOPV2 @ 6, 10, and 14 weeks 
265 
 
 
(2) B: nOPV2 + bOPV @ 6, 10, and 14 
weeks 
265 
 
 
(3) C: bOPV @ 6, 10, and 14 weeks 
265 
 
 
Total sample size 
795 
 
 
                                                                                                                                                  
Determination of Risk: Does the Research Involve (Check all that apply) 
  Human exposure to radioactive agents? 
  Foetal tissue or abortus? 
  Investigational new device? 
        Specify: __________________ 
  Existing data available from Co-investigator? 
 
 
  Human exposure to infectious agents? 
  Investigational new drug/vaccine? 
  Existing data available via public archives/sources? 
  Pathological or diagnostic clinical specimen only? 
  Observation of public behaviour? 
    
  New treatment regime? 
 
Will the information be recorded in such a manner that study participants can be identified from the 
information directly or through identifiers linked to the study participants? 
Yes 
 
No 
 
Does the research deal with sensitive aspects of the study participants’ sexual behaviour, alcohol use or 
illegal conduct such as drug use? 
Yes 
 
No 
 
 
  Could information on study participants, if available to people outside of the research team: 
 
a) Place them at risk of criminal or civil liability? 
Yes 
 
No 
 
b) Damage their financial standing, reputation or employability, or social rejection, or lead to 
stigma, divorce etc.? 
Yes 
 
No 
 
 
Do you consider this research: (check one) 
 
   Greater than minimal risk 
 No more than minimal risk 
   Only part of the diagnostic test 
 
Note: Minimal Risk: The probability and the magnitude of the anticipated harm or discomfort to participants is not greater than those ordinarily encountered 
in daily life or during the performance of routine physical, psychological examinations or tests, e.g. the risk of drawing a small amount of blood from a 
healthy individual for research purposes is no greater than when the same is performed for routine management of patients. 

 
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Risk Group of Infectious Agent and Use of Recombinant DNA 
 
a) Will specimens containing infectious agent be collected? 
 Yes            
 No            
 Not applicable 
b) Will the study involve amplification by culture of infectious 
agents? 
 Yes            
 No            
 Not applicable 
c) If response to questions (a) and/or (b) is ‘yes’, to which Risk 
Group (RG) does the agent(s) belong? (Please visit 
http://www.icddrb.net.bd/jahia/Jahia/pid/684 to review list of 
microorganism by Risk Group) 
 RG1           
 RG2         
 RG3           
 RG4 
d) Does the study involve experiments with recombinant DNA? 
 Yes            
 No            
 Not applicable 
Does the study involve any biohazards materials/agents or microorganisms of risk group 2, 3, or 4 (GR2, GR-3 or 
GR4)? 
 
 
 Yes            
 No            
 
[If the response is ‘yes’] I, (print name of the PI) affirm that we will use the standard icddr,b laboratory procedures 
for biosafety of the hazardous materials/agents or microorganisms in the conduction of the study.  
 
 
_________________________________                                                                                        _____________ 
Signature of  the Principal Investigator                                                                                                  Date 
Dissemination Plan: [please explicitly describe the plans for dissemination, including how the research findings would be 
shared with stakeholders, identifying them if known, and the mechanism to be used; anticipated type of publication (working 
papers, internal (institutional) publication, international publications, international conferences/seminars/workshops/ agencies. 
[Check all that are applicable] 
 
Dissemination type 
Response 
Description (if the response is a yes) 
Seminar for icddr,b 
scientists/ staff 
 No 
Yes 
The results will be shared in icddr,b internal seminars 
Internal publication 
No 
 Yes 
 
Working paper 
 No 
 Yes 
 
Sharing with GoB (e.g. 
DGHS/ Ministry, others) 
 No 
 Yes 
The clinical trials results could inform government of Bangladesh 
immunization policy 
Sharing with national 
NGOs 
 No 
 Yes 
The results will be shared with national NGOs 
Presentation at national 
workshop/ seminar 
 No 
 Yes 
The results will be presented at a national workshop/seminar 
Presentation at 
international workshop/ 
conference 
 No 
 Yes 
The results will be shared in international conferences as these results 
could have global implication 
Peer-reviewed publication 
 No 
Yes 
Multiple peer-reviewed publications are possible 
Sharing with international 
agencies 
 No 
 Yes 
The results will be shared with international agencies such as World 
Health Organization 
Sharing with donors 
 No 
 Yes 
CDC is funding agency for this clinical trial and the results will be shared 
with CDC 
Policy brief 
 No 
 Yes 
 
Other 
 
 
 
Other 
 
 
 

 
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` 
Funding: 
Is the protocol fully funded? 
 Yes 
 No 
If the answer is yes, please provide 
sponsor(s)’s name 
1. CDC through co-operative agreement.  
2. 
Is the protocol partially funded? 
 Yes 
 No 
If the answer is yes, please provide 
sponsor(s)’s name 
1. 
2. 
 
If fund has not been identified: 
Is the proposal being submitted for funding? 
 Yes 
 No 
If yes, name of the funding agency 
1. 
2. 
 
Conflict of interest: 
Do any of the participating investigators and/or member(s) of their immediate families have an equity relationship (e.g. 
stockholder) with the sponsor of the project or manufacturer and/or owner of the test product or device to be studied or serve 
as a consultant to any of the above? 
 No 
 Yes (please submit a written statement of disclosure to the Executive Director, icddr,b) 
Proposed Budget: 
Dates of Proposed Period of Support             
 
 
            Cost Required for the 
Budget Period ($)  
 
(Day, Month, Year - DD/MM/YY)                      
    
 Beginning Date: September 2020 
  
 End Date: June 2021 
 
 
 
 
 
 
Years 
Direct Cost 
Indirect 
Cost 
Total 
Cost 
Year-1 
586,896 
174,938 
761,834 
Year-2 
      
      
 
Year-3 
      
      
 
Year-4 
      
      
 
Year-5 
      
      
 
Total 
586,896 
174,938 
761,834 
 
Certification by the Principal Investigator: 
 
I certify that the statements herein are true, complete and accurate to the best of my knowledge. I am aware that any false, 
fictitious, or fraudulent statements or claims may subject me to criminal, civil, or administrative penalties. I agree to accept the 
responsibility for the scientific conduct of the project and to provide the required progress reports including updating protocol 
information in the NAVISION if a grant is awarded as a result of this application. 
 
I also certify that I have read icddr,b Data Policies and understand the PIs’ responsibilities related to archival and sharing of 
research data, and will remain fully compliant to the Policies. (Note: The Data Policies can be found here: 
http://www.icddrb.org/who-we-are/data-policies) 
 
 
___________                                                                                                                   ____________ 
Signature of PI                                                                                                                                         Date              
                                       

 
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Approval of the Project by the Division Director of the Applicant: 
 
 
The above-mentioned project has been discussed and reviewed at the Division level. 
 
 
 
                         
                 ___________ 
      
Name of the Division Director 
Signature 
                Date of Approval 
 
 
 

 
14 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Table of Contents 
 
RRC APPLICATION FORM .............................................................................................................. 1 
Protocol Version Control Page........................................................................................................... 15 
Glossary of Terms .............................................................................................................................. 16 
Project Summary ................................................................................................................................ 17 
Description of the Research Project ................................................................................................... 19 
Hypothesis to be tested: ..................................................................................................................... 19 
Background of the Project including Preliminary Observations: ...................................................... 21 
Research Design and Methods ........................................................................................................... 24 
Sample Size Calculation and Outcome Variables .............................................................................. 32 
Timeline of activities.......................................................................................................................... 34 
Data Analysis ..................................................................................................................................... 34 
Data Safety Monitoring Plan (DSMP) ............................................................................................... 37 
Ethical Assurance for Protection of Human rights ............................................................................ 40 
Use of Animals ................................................................................................................................... 43 
Collaborative Arrangements .............................................................................................................. 43 
Facilities Available............................................................................................................................. 44 
Literature Cited .................................................................................................................................. 44 
Budget ................................................................................................................................................ 46 
Budget Justifications .......................................................................................................................... 47 
Other Support ..................................................................................................................................... 48 
Biography of the Investigators ........................................................................................................... 49 
Consent (English and Bangla) ............................................................................................................ 66 
Information Sheet for Participant ....................................................................................................... 66 
Appendix A: Study Forms ................................................................................................................. 76 
Appendix B: Microneutralization Test for Polio Antibodies ........................................................... 103 
Appendix C: Summary of Laboratory Methods for Stool Testing .................................................. 115 
Appendix D: External Reviews ........................................................................................................ 117 
Check List…………………………………………………………………………………………..120 
E-mail approvals of Investigators…………………………………………………………………..121 
 
 
 
 
 
 
 
   
  Check here if appendix is included 
  
 
 
 
 
 
 
 
 
 
 

 
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Protocol Version Control Page 
 
Listed below is the guidance for protocol version control. The table below lists the protocol versions, date of 
activation of a version, the name of the study staff who finalized the said protocol version and the reason for 
creating the protocol version compared to a prior existing version. 
 
1. Version number:  
a. The first draft of the document would be 1.01 
b. Subsequent drafts will have an increase of 0.01 (e.g. 1.02, 1.03, 1.04) 
c. The file name and footer of every page on the protocol will include the name of the protocol and 
version number. For this protocol the file name and footer will read: “nOPV2 and bOPV Study 
ver 1.XX”   
d. Each version of the protocol after 1.01 should have two copies: version labeled “track changes” 
and version labeled “clean”. Track change version will highlight the changes in the named 
version compared to prior.  
e. A pdf file of protocol should follow the same naming convention as the protocol in word 
processing software 
2. Date of activation  
a. Date the protocol version number went into effect as the study protocol 
b. Activation date can be skipped for development versions of the protocol, i.e.  versions before 
sharing with co-investigators 
3. Responsible study staff 
a. The study staff who is primarily responsible for creating the version and sharing with other study 
team 
4. Summary reason for protocol version 
a. A reason for creating the version and its eventual use, e.g. version for submission to external peer 
review, version for submission to RRC. One does not need to list the detailed changes in the 
named version compared to the prior version as that can be noted by the track changes copy 
Version number 
Date of 
Activation 
Staff Finalizing the 
Version 
Summary reason 
Ver 1.01 
05-Mar-2020 
Abhijeet Anand / 
Amanda Wilkinson  
Draft protocol shared with 
co-investigators 
Ver 1.02 
17-Mar-2020 
Amanda Wilkinson 
Revisions based on co-
investigator comments 
Ver 1.03 
08-Jun-2020 
Amanda Wilkinson 
Removed study arms with 
nOPV2 candidate 2, added 
stool collection at 8 weeks, 
updated study site 
Ver 1.04 
14-Jul-2020 
Amanda Wilkinson 
Revisions based on RRC 
comments 
Ver 1.05 
10-Aug-2020 
Amanda Wilkinson  
Revisions based on ERC 
comments 
Ver 1.06 
03-Dec-2020 
Amanda Wilkinson / 
Jaymin Patel  
Adding household 
transmission component 
 
 
 
 
 
 
 
 
 
 

 
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Glossary of Terms 
  
AE 
Adverse Event 
bOPV 
Bivalent Oral Poliovirus Vaccine 
CDC 
Centers for Disease Control and Prevention 
CRF 
Case Report Form (paper)  
cVDPV 
Circulating Vaccine-Derived Poliovirus 
DSMB 
Data Safety Monitoring Board 
eCRF 
Case Report Form (electronic/tablet) 
EPI 
Expanded Program on Immunization  
ERC 
Ethical Review Committee 
fIPV 
Fractional IPV 
GPEI 
Global Polio Eradication Initiative 
icddr,b 
International Center for Diarrhoeal Disease Research, Bangladesh 
IPV 
Inactivated Poliovirus Vaccine 
IRB 
Institutional Review Board 
nOPV2 
Novel monovalent oral poliovirus vaccine type 2 
OPV 
Oral Poliovirus Vaccine 
PID 
Participant Identification Number 
PCR 
Polymerase Chain Reaction 
RRC 
Research Review Committee 
SAE 
Serious Adverse Event 
SAGE 
Strategic Advisory Group of Experts on Immunization 
SC 
Subcutaneous 
SOP 
Standard operating procedure 
tOPV 
Trivalent Oral Poliovirus Vaccine 
VAPP 
Vaccine-Associated Paralytic Poliomyelitis 
VDPV 
Vaccine-Derived Poliovirus 
WHO 
World Health Organization 
WPV 
Wild poliovirus 
 
 
 

 
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Project Summary 
[The summary, within a word limit of 300, should be stand alone and be fully understandable.] 
Principal Investigator: Dr. K. Zaman 
 
Immunogenicity of novel monovalent oral poliovirus vaccine type 2 (nOPV2) with and without 
bivalent OPV 
Proposed start date: December 2020                                                           
Estimated end date: September 2021 
Background:  
Wild poliovirus type 2 (WPV2) was declared eradicated in 2015 and type 2 oral poliovirus vaccine 
(OPV2) was withdrawn globally in May 2016. Monovalent OPV2 (mOPV2) is stockpiled for 
responding to circulating type 2 vaccine-derived poliovirus (cVDPV2) outbreaks. However, Sabin 
polioviruses in OPV can revert to neurovirulence and there is evidence that mOPV2 vaccines used to 
control cVDPV2 outbreaks have generated new VDPV2 emergences that have led to cVDPV2 
outbreaks. Novel OPV2 (nOPV2) candidates have been developed with improved genetic stability to 
reduce the risk of seeding new VDPVs. Findings from a phase 1 clinical trial in adults demonstrated the 
safety and immunogenicity of nOPV2. Preliminary results from phase 2 clinical trials in adults and 
children also suggest that the nOPV2 vaccines are safe and immunogenic.  
As of late 2019, nOPV2 candidate 1 was selected for Emergency Use Listing submission and 
commercial scale manufacturing. It is expected that nOPV2 would replace mOPV2 for responding to 
type 2 outbreaks. Outbreak response for cVDPV2 also offers the opportunity to close immunity gaps to 
polioviruses types 1 and 3. Furthermore, GPEI might have to respond to two poliovirus outbreaks in the 
same geography. For either scenario, it would be important to get data on the immunogenicity of co-
administered nOPV2 and bOPV, compared to either vaccine given alone. No study has yet assessed the 
immunogenicity of co-administered nOPV2 and bOPV. The proposed study further builds on the 
growing evidence around nOPV2 use by investigating household transmission and the potential 
development of intertypic vaccine-derived recombinants following the first study vaccination. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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Objectives: 
 
Primary 
1. To compare type 2 immunogenicity of two doses of nOPV2 with two doses of nOPV2 co-
administered with bOPV.  
2. To compare type 1 and 3 immunogenicity of two doses of bOPV with two doses of bOPV co-
administered with nOPV2. 
Secondary 
1. To compare type 2 immunogenicity of one or three doses of nOPV2 with the same number of 
doses of nOPV2 co-administered with bOPV. 
2. To compare type 1 and 3 immunogenicity of one or three doses of bOPV with the same number 
of doses of bOPV co-administered with nOPV2. 
3. To assess the presence of fecal viral shedding (as determined by real-time RT-PCR) over time 
following one dose of nOPV2, bOPV, or co-administered nOPV2 and bOPV.   
4. To estimate household transmission of nOPV2 by assessing the presence of fecal viral shedding 
(as determined by real-time RT-PCR) among siblings of vaccine recipients.  
5. To assess the occurrence and genomic characteristics of intertypic recombinants in stool 
specimens of vaccine recipients and siblings after one dose of nOPV2 given with or without 
bOPV.  
 
Methods: 
 
This is an open-label randomized clinical trial. Vaccine recipient participants will be enrolled and 
randomized at 6 weeks to one of three arms:  
 
A: nOPV2 @ 6, 10, and 14 weeks 
B: nOPV2 + bOPV @ 6, 10, and 14 weeks 
C: bOPV @ 6, 10, and 14 weeks 
 
For each vaccine recipient, a sibling below 10 years of age will be selected for participation as a sibling.  
 
Outcome measures/variables: 
Poliovirus antibody titers to types 1, 2 and 3 will be measured in sera extracted from vaccine recipient 
blood collected at 6, 10, 14, and 18 weeks. Antibody titers at 6 weeks of age will be the assumed starting 
point for the expected decline in maternal antibody assuming a half-life of 28 days. Shedding of 
poliovirus will be measured from stool specimens collected from vaccine recipients and one sibling 
each at 1, 2, and 4 weeks post-vaccination in Arm A and at 2 and 4 weeks post-vaccination in Arms B 
and C. Intertypic recombinants will be characterized in a subset of stool specimens collected from 
vaccine recipients and siblings.  
 

 
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Description of the Research Project 
 
Hypothesis to be tested: 
 
Does this research proposal involve testing of hypothesis: 
 No       
 Yes (describe below)  
 
Primary hypotheses to be tested are:  
 
1. Type 2 immune response after two doses of nOPV2 with bOPV is non-inferior to the immune 
response observed after two doses of nOPV2 given alone (10% non-inferiority margin).  
[Primary objective: 1] 
2. Type 1 and 3 immune responses after two doses of bOPV with nOPV2 are non-inferior to the 
immune responses observed after two doses of bOPV given alone (10% non-inferiority margin).  
[Primary objective: 2] 
Secondary hypotheses to be tested are:  
 
1. Type 2 immune responses after one or three doses of nOPV2 with bOPV are non-inferior to the 
immune responses observed after the same number of doses of nOPV2 given alone (10% non-
inferiority margin).  
[Secondary objective: 1] 
2. Type 1 and 3 immune responses after one or three doses of bOPV with nOPV2 are non-inferior 
to the immune responses observed after the same number of doses of bOPV given alone (10% 
non-inferiority margin).  
[Secondary objective: 2] 
 
Exploratory questions: 
 
1. What are the shedding rates among vaccine recipients following one dose of nOPV2, bOPV, or 
co-administered nOPV2 and bOPV? 
[Secondary objective: 3] 
2. What is the shedding rate among siblings of vaccine recipients following one dose of nOPV2?  
[Secondary objective: 4] 
3. What is the estimated household transmission rate following one dose of nOPV2?   
[Secondary objective: 4] 
4. What proportion of vaccine recipients and siblings have intertypic recombinants in stool 
following one dose of nOPV2 given with or without bOPV? What are the characteristics of the 
recombinants? 
[Secondary objective: 5] 
 
Specific Objectives: 
 
Table 1: Schematic representation of study arms.  
 

 
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Primary objectives* 
 
1. To compare type 2 immunogenicity of two doses of nOPV2 with two doses of nOPV2 co-
administered with bOPV.  
a. nOPV2 at 6 and 10 weeks (A) vs. nOPV2 + bOPV at 6 and 10 weeks (B) 
2. To compare type 1 and 3 immunogenicity of two doses of bOPV with two doses of bOPV co-
administered with nOPV2.  
a. bOPV at 6 and 10 weeks (C) vs. nOPV2 + bOPV at 6 and 10 weeks (B) 
Secondary objectives* 
 
1. To compare type 2 immunogenicity of one or three doses of nOPV2 with the same number of 
doses of nOPV2 co-administered with bOPV. 
a. nOPV2 at 6 weeks (A) vs. nOPV2 + bOPV at 6 weeks (B) 
b. nOPV2 at 6, 10, and 14 weeks (A) vs. nOPV2 + bOPV at 6, 10, and 14  weeks (B) 
2. To compare type 1 and 3 immunogenicity of one or three doses of bOPV with the same number 
of doses of bOPV co-administered with nOPV2. 
a. bOPV at 6 weeks (C) vs. nOPV2 + bOPV at 6 weeks (B) 
b. bOPV at 6, 10, and 14 weeks (C) vs. nOPV2 + bOPV at 6, 10, and 14 weeks (B) 
3. To assess the presence of fecal viral shedding (as determined by real-time RT-PCR) over time 
following one dose of nOPV2, bOPV, or co-administered nOPV2 and bOPV.   
4. To estimate household transmission of nOPV2 by assessing the presence of fecal viral shedding 
(as determined by real-time RT-PCR) among siblings of vaccine recipients.  
5. To assess the occurrence and genomic characteristics of intertypic recombinants in stool 
specimens of vaccine recipients and their siblings after one dose of nOPV2 given with or 
without bOPV.  
 
*Note: For all objectives, timepoints refer to the timing of vaccine administration.  

 
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Background of the Project including Preliminary Observations: 
 
 
Polioviruses are a member of family Picornaviridae in the enterovirus subgroup, and poliomyelitis is an 
acute infection caused by polioviruses. Polioviruses are transmitted from person to person either faeco-
orally or through pharyngeal secretions. There is minimal cross-immunity between the three distinct 
serotypes of polioviruses (types 1, 2 and 3). After entering the body through the mouth, polioviruses 
replicate in the gastrointestinal tract for several days or weeks. In the majority of cases, they cause no 
apparent illness or mild illness including diarrhea, fever, and/or vomiting. In approximately one out of 
200 infections, however, polioviruses invade the cells of the spinal cord leading to paralysis of limb(s) 
or invade brain stem cells affecting the respiratory muscles. Of those individuals with paralytic 
poliomyelitis, an estimated 5-10% die because of breathing failure. The remaining suffer from lifelong 
paralysis of one or more limbs. Before the availability of vaccines, poliomyelitis was the leading cause 
of permanent disability worldwide [1, 2]. 
 
Poliovirus Vaccines 
Inactivated poliovirus vaccine (IPV) was licensed in 1955 in the United States and was the first licensed 
poliovirus vaccine. Subsequently, three type-specific, live, attenuated oral poliovirus vaccines were 
licensed in 1961–62. These monovalent oral poliovirus vaccines (mOPV) were quickly followed by 
licensure of trivalent OPV (tOPV) in 1963, which protects against all three poliovirus serotypes. 
Because of its low cost, ease of administration, induction of mucosal immunity, and ability to indirectly 
vaccinate susceptible contacts, tOPV largely replaced IPV as the vaccine of choice for polio vaccination 
in most countries [2-4].  
 
Despite their many advantages, there are important disadvantages with OPV use. Because Sabin viruses 
are live attenuated viruses, they can mutate during replication and revert to a neurovirulent form leading 
to vaccine-associated paralytic polio (VAPP). It is estimated a VAPP case occurs for every 1.2–1.5 
million OPV doses administered [2]. In addition, Sabin viruses can circulate in areas with inadequate 
polio vaccination coverage. Prolonged person-to-person transmission facilitates mutations and the 
emergence of virus variants called circulating vaccine-derived polioviruses (cVDPVs) which have re-
acquired the neurovirulence and transmissibility of wild poliovirus (WPV) [5]. Therefore, polio 
eradication will require eventual cessation of all traditional Sabin OPVs [6, 7].   
 
Immunological response after poliovirus vaccines is determined by measuring type-specific poliovirus 
antibodies using neutralization assays. After the first exposure to poliovirus antigens, which could be 
wild or vaccine-derived, a rapid rise in IgM titers is observed. This is followed by a slower rise in IgG 
titers, which typically peaks in four weeks [8]. 
 
OPV Cessation 
Wild poliovirus type 2 (WPV2) was declared eradicated in September 2015 [9, 10]. Since then, 
paralysis associated with type 2 poliovirus has continued mainly due to VDPVs from type 2 OPV. Due 
to the continued threat of paralysis from a mutated, neurovirulent and vaccine-derived type 2 poliovirus, 
the Strategic Advisory Group of Experts on Immunization (SAGE), a global advisory committee on 
immunization, recommended a phased cessation of OPV starting with type 2 oral poliovirus vaccine 
(OPV2) [11].  
 
Subsequently, OPV2 was withdrawn globally, including the replacement of tOPV with bivalent OPV 
(bOPV), containing poliovirus types 1 and 3, in May 2016. Since then, monovalent type 2 OPV (mOPV2) 
has been used for responding to circulating type 2 vaccine-derived poliovirus (cVDPV2) outbreaks. Since 
2016, over 300 million doses of mOPV2 have been used to respond to cVDPV2 outbreaks and some of 
these responses have led to emergence of new cVDPV2s, likely from the mOPV2 used during the 

 
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outbreak response. Therefore, novel OPV2 (nOPV2) candidates have been developed with improved 
genetic stability, thereby reducing the possibility of generating new VDPVs and cVDPV outbreaks [12, 
13].  
 
nOPV2 Candidates and Clinical Trials 
GPEI has been pursuing two candidate nOPV2 vaccines. To develop the current nOPV2 candidates, the 
existing OPV type 2 was modified to improve its genetic stability (additional details provided in the 
“Description of Study Vaccines” section of the protocol) [12-14]. For both nOPV2 candidates, clinical 
trials lots underwent standard monkey neurovirulence testing and the vaccine formulation testing used 
by Bio Farma for Sabin-based OPV products. 
 
Human clinical trials of nOPV2 began in 2017. As of June 2020, three human trials of nOPV2 have 
been conducted. 
 
The first human study was a double-blind, phase 1 trial conducted under biological containment in 
Belgium by the University of Antwerp. Thirty IPV-primed adults were randomized to receive one dose 
of either nOPV2 vaccine candidate. Both candidates were well tolerated; reversion to neurovirulence 
was low in shed virus when tested in transgenic mice, and >80% of participants demonstrated a type 2 
immune response [14].  
 
Phase 2 studies were implemented in Belgium and Panama to evaluate the safety and immunogenicity of 
both nOPV2 candidates. The University of Antwerp conducted a double-blinded, randomized, placebo-
controlled trial without containment that was completed in Q2 2019. The study enrolled 200 healthy OPV-
primed adults and 50 healthy IPV-only-primed adults and adolescents [15]. Preliminary results from the 
Belgium phase 2 trial suggest that both nOPV2 candidates (dose = 106 CCID50) are immunogenic. One-
dose seroconversion rates were ~75% for candidate 1 and ~51% for candidate 2, and both higher than the  
historical Sabin 2 control at a lower dose (29%; dose = 105 CCID50) (University of Antwerp, unpublished).  
 
In Panama, a phase 2 trial was implemented using an age de-escalation scheme. Participants were 100 
children aged 1–5 years with prior IPV and/or OPV vaccination in the first year of life, and 648 healthy 
infants who received bOPV (3 doses) and IPV (1 dose) prior to vaccination with nOPV2 at 18–22 weeks 
of age [16] . The infant component of the trial included both high-dose (106 CCID50) and low-dose (105 
CCID50) nOPV2 formulations. The main study activities were completed in Q3 2019, except for the 
longer-term safety follow up, which ended in early 2020.  Early results from the phase 2 trials in Belgium 
and Panama were reviewed by an independent data safety monitoring board, which supported age group 
progression from adults to toddlers and infants based on the available safety information. 
 
Toddlers in the Panama trial all received high-dose nOPV2; based on preliminary findings, one-dose 
seroconversion rates were high (~95%) for both candidates and exceed that of 105 CCID50 Sabin 2 (~63%). 
Among infants, immunogenicity non-inferiority to mOPV2 was achieved for both high- and low-dose 
candidate 1, but only for high-dose candidate 2 (A. Bandyopadhyay, unpublished). Both candidates were 
shown to replicate in the gut. Based on preliminary neurovirulence results, both candidates appear to be 
more phenotypically stable than Sabin 2; however, candidate 1 performed better than candidate 2, i.e. had 
significantly lower rates of paralysis in the mouse model. There was no evidence of viral recombination 
in the sequencing results (A. Bandyopadhyay, unpublished).  
 
Overall, trial results to date suggest that nOPV2 is safe, immunogenic, and more genetically stable than 
Sabin 2 [16]. As of late 2019, nOPV2 candidate 1 was selected for Emergency Use Listing submission 
and commercial scale manufacturing.  
 

 
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Two upcoming trials of nOPV2 candidate 1 are planned. In Bangladesh, icddr,b will implement a phase 
2 study on the safety and immunogenicity of nOPV2 in OPV-naïve newborns. A phase 3 trial will be 
conducted in The Gambia to assess expanded safety and lot-to-lot consistency of nOPV2. 
 
Intertypic Recombination 
Enteroviruses, such as poliovirus, have a large degree of genetic variability and evolve through mutation 
and recombination. Recombination can occur between different Sabin vaccine strains, or Sabin virus 
can recombine with wild polio strains or non-polio enteroviruses [17]. For recombination to occur, a 
cell needs to be simultaneously infected with two different viral strains. Recombination is a common 
event among OPV recipients and can occur soon after vaccination; it rarely results in negative outcomes 
such as VAPP [18-22]. Intertypic recombination is thought to contribute to VDPV emergence as even a 
single recombination event can lead to replacement and reversion of attenuating elements. The nOPV2 
candidates were designed to be more genetically stable than mOPV2; however, their recombination 
behaviour in settings with high prevalence of enterovirus infection or other OPV strains is still 
unknown.  
 
Justification for this Study 
It is expected that nOPV2 would replace mOPV2 for responding to type 2 outbreaks when authorized by 
Emergency Use Listing and/or licensed and available for use. Vaccination campaigns as part of outbreak 
response for cVDPV2 also offers the opportunity to close immunity gaps to polioviruses types 1 and 3 
through co-administration of bOPV. Furthermore, GPEI might have to respond to two poliovirus 
outbreaks in the same geography such as WPV1 and cVDPV2 in Nigeria, Pakistan in 2016, and 
Afghanistan and Pakistan in 2019–2020; cVDPV2 and cVDPV3 in Somalia in 2018; and cVDPV2 and 
cVDPV1 in the Philippines and Malaysia in 2019/2020. It would be important to get data on 
coadministration of nOPV2 and bOPV, particularly to note if there is any inhibitory effect on the 
immunogenicity of either vaccine.  
 
No study has assessed co-administration of nOPV2 and bOPV; therefore, the study we have proposed 
compares the type-specific immunogenicity of co-administered nOPV2 and bOPV with each vaccine 
given alone. This clinical trial will build on existing evidence for nOPV2 immunogenicity and generate 
data to inform global poliovirus outbreak response strategies.  
 
Although nOPV2 was designed to be more genetically stable than mOPV2, there is a lack of real-world 
evidence on its recombination behaviours. Therefore, in the current study, we will assess recombinants 
at two timepoints for all arms (8 and 10 weeks; two and four weeks post-vaccination, respectively). 
Timepoints for stool collection were selected based on when shedding would be expected to be high in 
children without pre-existing mucosal immunity.  
 
While phase I and II trials have measured shedding post-vaccination among nOPV2 vaccine recipients, 
these were among individuals with previous poliovirus vaccination [14, 16]. There is also currently a 
lack of data to estimate potential secondary transmission of the vaccine virus to close contacts of 
vaccine recipients. The household transmission component of the study will be descriptive of not 
include a Sabin 2 comparison group; nevertheless, it is important to assess the transmissibility of 
nOPV2 in order to better anticipate risks. 
 
Overall, these data will be programmatically valuable and can inform future decision making around 
nOPV2 use, particularly for outbreak response.  
 

 
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Research Design and Methods 
Study Area 
Mirpur is one of the 14 Thanas of Dhaka city with a population of about two million in an area of 59 
square kilometers.  Mirpur Thana of Dhaka city is divided into 14 sections. The area is densely 
populated and is about eight kilometers from icddr,b’s Dhaka Hospital. The population is stable with 
low socioeconomic conditions.  The average income in the slum areas of Mirpur is Tk.4200 (about US 
$55) per month per family. Only 25% of fathers and 15% of mothers have more than five years of 
formal education. The study will be conducted in sections 1- 2, 6-7 and 10-13 of Mirpur, which has a 
population of about 500,000. icddr,b and CDC have collaborated on multiple polio vaccine trials that 
were conducted in the study clinic in Mirpur [23, 24].  
 
If feasible based on guidelines/recommendations related to the COVID-19 pandemic (e.g., physical 
distancing), the study will also be conducted at the clinical trial unit (CTU), Dhaka. The study area 
includes the adjoining slum and non-slum areas of icddr,b Mohakahli, Dhaka - Kunipra, Arjat Para, 
Nakhalpara, Korail, Begunbari, Chairman bari and Badda. Participants will attend CTU, which is about 
one kilometer from the areas. The population is approximately 300,000 and literacy is ~50%. It is stable 
with low socio-economic conditions. The population is mainly engaged in industrial labour, garment 
factory, small businesses and rickshaw pullers.  
 
Study staff will identify infants eligible to be vaccine recipients through house-to-house visits at the two 
proposed sites of Mirpur and Mohakahli.  
 
Study Design 
This is an open-label, non-inferiority, balanced three arm randomized clinical trial assessing 
immunogenicity of poliovirus vaccines. Vaccine recipients will be identified through active surveillance 
of new births in the community and parents will be requested to participate through home visits by local 
field workers. Vaccine recipients will be enrolled at 6 weeks of age, randomly assigned to one of the three 
arms and followed to 18 weeks of age.  
 
Inclusion Criteria  
Criteria for both vaccine recipient and sibling must be met for enrolment in the trial.  
Vaccine Recipient 
1. Healthy infants 6 weeks of age (range: 42–48 days).   
2. Parents that consent for participation in the full length of the study.   
3. Parents that can understand and comply with planned study procedures. 
4. Infant has at least one sibling aged <10 years living in the same household that is eligible for 
participation in the study and can be enrolled at the same time as the vaccine recipient.  
Sibling 
1. Healthy child aged <10 years.  
2. Sibling of the vaccine recipient and resides in the same household.  
3. Parents that consent for participation in the study until and including the Clinic Visit at 10 weeks 
(age of vaccine recipient). 
4. Parents that can understand and comply with planned study procedures 
 
Exclusion Criteria 
If either vaccine recipient or sibling meets the exclusion criteria, then neither can be enrolled in the 
trial. 

 
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Vaccine Recipient 
1. Parents, infants, and siblings who are unable to participate in the full length of the study (e.g., 
plan to move away from the study area during the study period). 
2. A diagnosis or suspicion of immunodeficiency disorder either in the infant or in an immediate 
family member. 
3. A diagnosis or suspicion of bleeding disorder that would contraindicate administration of bOPV 
or nOPV2 or collection of blood by venipuncture. 
4. Acute diarrhoea, infection or illness at the time of enrolment (6 weeks of age) that would require 
infant’s admission to a hospital. 
5. Acute vomiting and intolerance to liquids within 24 hours before the enrolment visit (6 weeks of 
age).  
6. Evidence of a chronic medical condition identified by a study medical officer during physical 
exam. 
7. Receipt of any polio vaccine (OPV or IPV) before enrolment based upon documentation or 
parental recall. 
8. Known allergy/sensitivity or reaction to polio vaccine, or its contents. 
9. Infants from multiple births. Infants from multiple births will be excluded because the infant(s) 
who is/are not enrolled would likely receive OPV through routine immunization and transmit 
vaccine poliovirus to the enrolled infant. Even if all births from a multiple birth could be 
enrolled in the study, we will exclude multiple births as discontinuation of one may lead to 
discontinuation of multiple vaccine recipients. 
10. Infants from premature births (<37 weeks of gestation). 
Sibling 
1. Participant in a previous poliovirus vaccine clinical trial.  
2. Unable to participate in the study until and including the Clinic Visit at 10 weeks (age of vaccine 
recipient). 
3. A diagnosis or suspicion of immunodeficiency disorder either in the sibling or in an immediate 
family member. 
4. Acute diarrhoea, infection or illness at the time of enrolment that would require sibling’s 
admission to a hospital. 
5. Evidence of a chronic medical condition identified by a study medical officer during physical 
exam.  
 
Discontinuation Criteria 
Vaccine Recipient 
1. Withdrawal of consent for participation for any reason. 
2. Request by parents of vaccine recipient to terminate all study procedures. 
3. Identification of immunodeficiency disorder, bleeding disorder or another medical condition for 
which continued participation, in the opinion of the principal investigator, would pose a risk to 
the vaccine recipient to continue in the study. 
4. Receipt of immunosuppressive medications. 
5. Receipt of any polio (OPV or IPV) vaccine outside of study after enrolment (as per parent’s 
report). Vaccine recipients who received polio vaccines outside of the study, including any polio 
vaccine administered in polio campaigns, would be excluded from further study procedures. 
6. Temporary discontinuation may occur when there has been a temporary suspension of study 
activities. Study enrolment will be suspended from 12 weeks before the start of polio campaigns 
in the study areas to 6 weeks after the completion of polio campaigns in the study areas. This 
measure is to prevent potential interference from polio vaccine administered during polio 
campaigns in the study areas. If vaccine recipients receive vaccine during polio campaigns, they 
would be discontinued from further study procedures. 

 
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7. Unable to collect or obtain blood at enrolment (Clinic Visit at 6 Weeks of Age). 
8. Allergic reaction to a prior dose of polio vaccine, or its contents. 
9. Premature termination of the study. 
Sibling 
1. Vaccine recipient is discontinued from the study. 
2. Withdrawal of consent for participation for any reason. 
3. Receipt of any polio (OPV or IPV) vaccine after enrolment (as per parent’s report). Siblings 
who receive polio vaccines during their participation in the study, including any polio vaccine 
administered in polio campaigns, would be excluded from further study procedures. 
 
Description of Study Vaccines 
Two polio vaccines will be used in the study: bOPV and nOPV2  
 
1. Bivalent oral polio vaccine (bOPV) 
a. Manufacturer: Bio Farma 
b. Presentation: the vaccine will be available in multi-dose vials 
c. Formulation: Each dose (2 drops = 0.1 ml). 
d. Administration: one vaccine vial will be used per participant. 
e. Previous use at icddr,b: bOPV is part of the routine immunization schedule in Bangladesh.  
 
2. Candidate 1 novel monovalent oral poliovirus vaccine type 2 (nOPV2)  
a. Manufacturer: Bio Farma 
b. Description: nOPV2 candidate 1 (S2/cre5/S15domV/rec1/hifi3) is a live-attenuated serotype-
2 poliovirus that was derived from a modified Sabin type-2 infectious cDNA clone and 
propagated in Vero cells. To improve genetic stability, nucleotide sequence modifications 
were made in the major determinant for attenuation in the Sabin 5’-untranslated region. 
Additionally, two modifications in the polymerase 3D were made to further improve stability 
of the attenuation, and a key replication element from the 2C coding region was relocated to 
the 5ʹ-untranslated region to inhibit recombination [14].  
c. Manufacturer: Bio Farma 
d. Presentation: the vaccine will be available in multi-dose vials.  
e. Formulation: approximately 105 CCID50/dose, dose = 2 drops 
f. Administration: one vaccine vial will be used per participant. 
g. Previous use at icddr,b: Prior to the start of the current study, icddr,b will implement a naïve 
infants trial that includes nOPV2 (PR-20001). 
 
Disposal of Used Study Vaccines 
At the end of each day, the used study vaccine vials will be returned from the study clinics to icddr,b 
where they will be stored. After using all doses in the vial or after having passed the usage duration 
recommended by the manufacturer, the vaccine vials will be stored for the duration of the study but not 
used for administration to participants. This will be done to ensure availability of the used vaccine vials 
should a need arise, such as an investigation of adverse events. The used vaccine vials will be stored at 
least three months after the completion of data analysis for the study. Four weeks prior to the planned 
disposal of the used vaccine vials, the PI will notify the Data Safety and Monitoring Board (DSMB) of 
the intent to destroy the used vaccine vials by a specific date; this may be delayed at the request of 
DSMB. Unless the DSMB requests the PI to delay disposal of the used vaccine vials, the used vaccine 
vials will be destroyed per icddr,b’s standard procedures for biohazardous materials. 

 
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Study Procedures 
Field workers will inform parents about the study and briefly explain the study procedures, including 
stool sampling from both the vaccine recipient and the sibling. If interested in participating, parents will 
be asked to visit the study clinic when the vaccine recipient is 6 weeks of age to obtain comprehensive 
information on the study and to complete enrolment procedures; both the vaccine recipient and sibling  
will need to be present at this visit. Parents will be provided transport costs for all study visits to/from 
the clinic.  
 
The below description summarizes the study clinic and home visits over the course of the study.  
Vaccine recipients may additionally visit the study clinic for routine childhood vaccinations or for 
management of adverse events. Although non-study vaccines included in the routine immunization 
schedule in Bangladesh will be available and vaccination will be encouraged, parents will have the final 
decision to vaccinate their infant. Injectable non-polio, routine immunization vaccines will be 
administered at the recommended body site per the national guidelines of Government of Bangladesh. If 
a study visit involves collection of blood as well as administration of study vaccines, then blood 
collection will precede study vaccine administration. For Arm B, concomitant administration will be 
defined as nOPV2 provided first, followed by bOPV on the same day in rapid succession with a target 
of ≤5 minutes apart.   
 
Enrolment visit – 6 weeks of age (acceptable range 42–48 days) [All arms; sibling must also be 
present at this clinic visit, but not subsequent clinic visits] 
During this visit, the following procedures will be conducted: 
1. The medical officer will confirm that the vaccine recipient and sibling are eligible to participate. If 
the vaccine recipient and sibling are eligible, all study procedures will be explained to the parents. 
The parents will be requested to provide informed consent; consent will be documented per study 
procedures.  
2. The vaccine recipient will be assigned a participant identification number (PID) and to a study arm 
according to a pre-determined randomization scheme. Further details of the randomization scheme 
are described in the “Data Analysis” section. 
3. Vaccination record of the sibling will be obtained and recorded by the medical officer. 
4. Socio-demographic and clinical information will be recorded on the corresponding electronic case 
report form (eCRF).  
5. One blood specimen (1 ml) will be collected from the vaccine recipient by venipuncture. 
6. The first dose of study vaccines will be administered to the vaccine recipient, as directed by study 
arm.   
7. The vaccine recipient will be observed for 30 minutes to monitor for any adverse reactions from the 
blood collection or administration of study vaccines. 
8. The vaccine recipient will receive the recommended routine childhood vaccinations, except polio 
vaccination. Routine polio vaccines will be replaced by the study vaccine.   
9. Clinical information will be collected about the sibling. The sibling must be present at this visit so 
that he/she may be examined by a medical officer but does not need to be present for any subsequent 
clinic visits.  
10. Schedule the household visit(s) and the next clinic visit. 
 
Phone call or home Visit (24–48 hours after the first study vaccination) [All arms] 
1. A study staff will contact the parents of the vaccine recipient to inquire and record any potential 
adverse events after the first study vaccination.  
2. This phone call or visit will be conducted 24–48 hours after the first study vaccination visit. 
 

 
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Home Visit – 7 weeks of age [Arm A only]   
Home Visit #1 (two days prior to 7 weeks of age) 
1. The study field worker will visit the vaccine recipient’s home and provide two plastic screw-cap stool 
containers to the mother. She will review the stool collection procedures with the mother, including 
sibling sampling.  
2. The study field worker will complete the top portion of the study questionnaire. 
 
Home Visit #2 (7 weeks of age)  
1. Mother will collect stool specimens from the vaccine recipient and sibling. If possible, mother will 
collect fresh stool from each child’s diaper or have the child defecate into a clean plastic pot. Mother 
will be asked to collect stool about the size of one adult thumb (8 grams).  
She will place the containers with the collected stool specimens immediately into the freezer (if 
available) or in a cool place in the home. The same procedures will be used for collecting stool 
specimens from the sibling. Mother will immediately notify the study field worker after each stool 
collection.    
2. Within two hours of notification, the study field worker will go to the vaccine recipient’s home with 
ice packs, two stool containers, and a stool carrier bag. If necessary, she will transfer stool from the 
original stool container to a new stool container. Otherwise, the study field worker will place the stool 
containers in the stool carrier bag, between the two ice packs.  
3. The study field worker will complete the rest of the study questionnaire. 
4. The study field worker will transport the stool samples to the study clinic within 30 minutes of pick 
up and store the stool containers at 2-8oC. 
5. The study field worker will provide the completed study questionnaire to the data entry staff. 
6. Data entry staff will enter the completed study questionnaire into REDCap (eCRF).  
 
 
Home Visit – 8 weeks of age [All arms]   
Home Visit #1 (two days prior to 8 weeks of age) 
1. The study field worker will visit the vaccine recipient’s home and provide two plastic screw-cap 
stool containers to the mother. She will review the stool collection procedures with the mother, 
including sibling sampling.  
2. The study field worker will complete the top portion of the study questionnaire. 
 
Home Visit #2 (8 weeks of age)  
1. Mother will collect stool specimens from the vaccine recipient and sibling. If possible, mother will 
collect fresh stool from each child’s diaper or have the child defecate into a clean plastic pot. Mother 
will be asked to collect stool about the size of one adult thumb (8 grams).  
She will place the containers with the collected stool specimens immediately into the freezer (if 
available) or in a cool place in the home. The same procedures will be used for collecting stool 
specimens from the sibling. Mother will immediately notify the study field worker after each stool 
collection.    
2. Within two hours of notification, the study field worker will go to the vaccine recipient’s home with 
ice packs, two stool containers, and a stool carrier bag. If necessary, she will transfer stool from an 
original stool container to a new stool container. Otherwise, the study field worker will place the 
stool containers in the stool carrier bag, between the two ice packs.  
3. The study field worker will complete the rest of the study questionnaire. 
4. The study field worker will transport the stool samples to the study clinic within 30 minutes of pick 
up and store the stool containers at 2-8oC. 
5. The study field worker will provide the completed study questionnaire to the data entry staff. 
6. Data entry staff will enter the completed study questionnaire into REDCap (eCRF).  

 
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Home Visit – 10 weeks of age [All arms]  
Home Visit #1 (two days prior to 10 weeks of age) 
1. The study field worker will visit the vaccine recipient’s home and provide two plastic screw-cap 
stool containers to the mother. She will review the stool collection procedures with the mother, 
including sibling sampling.  
2. The study field worker will complete the top portion of the study questionnaire. 
 
Home Visit #2 (10 weeks of age)  
1. Mother will collect stool specimens from the vaccine recipient and sibling. If possible, mother will 
collect fresh stool from each child’s diaper or have the child defecate into a clean plastic pot. Mother 
will be asked to collect stool about the size of one adult thumb (8 grams).  
She will place the containers with the collected stool specimens immediately into the freezer (if 
available) or in a cool place in the home. The same procedures will be used for collecting stool 
specimens from the sibling. Mother will immediately notify the study field worker after each stool 
collection.    
2. Within two hours of notification, the study field worker will go to the vaccine recipient’s home with 
ice packs, two stool containers, and a stool carrier bag. If necessary, she will transfer stool from an 
original stool container to a new stool container. Otherwise, the study field worker will place the 
stool containers in the stool carrier bag, between the two ice packs.  
3. The study field worker will complete the rest of the study questionnaire. 
4. The study field worker will transport the stool samples to the study clinic within 30 minutes of pick 
up and store the stool containers at 2-8oC. 
5. The study field worker will provide the completed study questionnaire to the data entry staff. 
6. Data entry staff will enter the completed study questionnaire into REDCap (eCRF).  
 
Study Clinic Visit – 10 weeks of age [All arms]  
During this visit, the following procedures will be conducted: 
1. The medical officer will complete the eCRF and examine the vaccine recipient.  
2. One blood specimen (1 ml) will be collected by venipuncture. 
3. Study vaccines will be administered, as directed by study arm.  
4. The vaccine recipient will be observed for 30 minutes to monitor for any adverse reactions to the 
study vaccine. 
5. The vaccine recipient will receive the recommended routine childhood vaccinations, except polio 
vaccination. Routine polio vaccines will be replaced by the study vaccine. 
6. Schedule the next visit. 
 
Study Clinic Visit – 14 weeks of age [All arms]  
During this visit, the following procedures will be conducted: 
1. The medical officer will complete the eCRF and examine the vaccine recipient.  
2. One blood specimen (1 ml) will be collected by venipuncture. 
3. Study vaccines will be administered, as directed by study arm.  
4. The vaccine recipient will be observed for 30 minutes to monitor for any adverse reactions to the 
study vaccine. 
5. The vaccine recipient will receive the recommended routine childhood vaccinations, except polio 
vaccination. Routine polio vaccines will be replaced by the study vaccine. 
6. Schedule the next visit. 
 
Study Clinic Visit – 18 weeks of age [All arms] 

 
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During this visit, the following procedures will be conducted: 
1. The medical officer will complete the eCRF and examine the vaccine recipient.  
2. One blood specimen (1 ml) will be collected by venipuncture. 
3. Completion of study activities.  
4. Vaccine recipients in Arms A and B will be given the first of three bOPV doses. Note: Arm B will 
be given three doses of bOPV to account for possible immunologic interference during co-
administration of bOPV with nOPV2. All vaccine recipients will be given the first of two fIPV 
doses.  
Timing of Evaluations 
Entry/eligibility evaluations 
• Entry evaluations will be completed at 6 weeks of age. 
• Blood collection and study vaccine administration are planned during the study entry evaluation at 6 
weeks of age. Blood will be collected before administration of any study vaccines. On the date of 
enrolment, study vaccines will be administered only if adequate blood specimen has been collected. 
If blood cannot be collected on the day of enrolment, study vaccine recipients will be discontinued 
from the study.  
• The window for entry evaluation including collection of blood and administration of vaccine is 42–
48 days of age.   
 
Post-entry evaluations and follow-up 
• After the first enrolment visit (6 weeks of age, range 42–48 days), the window for study visits is +/- 
3 days. 
• Blood will be collected before any study or EPI vaccines are administered. 
• Stool collection from vaccine recipients and one sibling each will take place at 8 and 10 weeks of 
age for all Arms, and additionally at 7 weeks for Arm A only.  
 
Concomitant Medications and Vaccines 
There will be no restrictions on the use of medications or treatments for concomitant diseases. If the 
vaccine recipient needs to receive medications that cause immunosuppression, the vaccine recipient will 
be discontinued from the study.  
 
If the vaccine recipient receives a dose of either polio vaccines (OPV or IPV) outside of the study 
during the study period, the vaccine recipient will be discontinued from the study. Parents will be 
encouraged and offered an opportunity to have their child vaccinated with routine childhood 
immunizations during study clinic visits, in accordance with Bangladesh EPI’s vaccination schedule. 
Other non-routine immunizations will not be prohibited if they do not contain polio vaccines.   
 
Specimen Collection, Handling and Testing 
Sera: Table 2 summarizes the blood samples that will be collected for serological testing. All study 
visits requiring blood collection will collect 1 ml of blood. Blood will be stored temporarily in the study 
clinic and transported from the clinic to the icddr,b virology laboratory at 2–8 degrees Celsius within 6 
to 8 hours of collection. Then sera will be separated by centrifugation in icddr,b virology lab within one 
hour. Tubes with haemolysed blood will be discarded following standard biohazard waste disposal 
protocol. After sera have been aliquoted separately for testing of poliovirus (0.2 mL), the remaining sera 
will be stored at icddr,b in cryovials with CDC provided labels that include vaccine recipient 
identification number (PID) and study protocol number. At all visits, only one aliquot (0.2 mL) will be 
prepared for shipment for Atlanta. The samples stored at icddr,b are reserve samples that may need to 
be shipped to CDC in the event that samples in the initial shipment were adversely affected, resulting in 

 
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the inability to perform testing or uninterpretable results. Cryovial tubes that will be sent to CDC for 
testing will be stored in a -20 degree Celsius freezer. The rest of the aliquoted sera will be stored at -70 
degree Celsius at icddr,b. 
 
Table 2. Summary of sera samples for serological testing 
 
Age of vaccine 
recipient 
Blood  
collected from 
arms 
Volume of 
blood 
collection 
Sera available  Sera for 
poliovirus 
testing 
Sera stored at 
icddr,b 
6 weeks 
All arms 
1 ml 
0.4 ml 
0.2 ml 
0.2 ml 
10 weeks 
All arms 
1 ml 
0.4 ml 
0.2 ml 
0.2 ml 
14 weeks 
All arms 
1 ml 
0.4 ml 
0.2 ml 
0.2 ml 
18 weeks 
All arms 
1 ml 
0.4 ml 
0.2 ml 
0.2 ml 
 
Stool: For the follow-ups at 7 (Arm A only), 8, and 10 weeks of age, parents of enrolled participants 
will be requested to collect about 8 grams (about the size of one adult thumb) of stool from vaccine 
recipients and from one sibling each. After collection, specimens will be kept at a temperature of 2–8 
degrees Celsius at the study clinic and transferred to icddr,b within the same day. Upon arrival at 
icddr,b, samples will be aliquoted and stored at -20 degrees Celsius. Two aliquots will be prepared for 
each stool sample; 4 g will be aliquoted for shipment and testing and a second aliquot will be stored at 
icddr,b. Aliquots will be labelled with CDC-provided labels that include PID, indication of whether the 
sample corresponds to a vaccine recipient or sibling, and date of collection. 
 
Table 3. Summary of stool samples for testing 
 
Age of vaccine 
recipient 
Stool collected 
from arms 
Volume of 
stool collection 
Stool for 
testing 
Stool stored at icddr,b 
7 weeks 
Vaccine 
recipient: 
Arm A only 
Approximately 
8 g 
4 g 
Remaining stool quantity (no 
more than 4 grams) 
Sibling:  
One sibling 
from Arm A 
only 
Approximately 
8 g 
4 g 
Remaining stool quantity (no 
more than 4 grams) 
8 weeks 
Vaccine 
recipient: 
All arms 
Approximately 
8 g 
4 g 
Remaining stool quantity (no 
more than 4 grams) 
Sibling: 
One sibling 
from all arms 
Approximately 
8 g 
4 g 
Remaining stool quantity (no 
more than 4 grams) 
10 weeks 
Vaccine 
recipient: 
All arms 
Approximately 
8 g 
4 g 
Remaining stool quantity (no 
more than 4 grams) 
Sibling: 
One sibling 
from all arms 
Approximately 
8 g 
4 g 
Remaining stool quantity (no 
more than 4 grams) 
 
Sera and stool specimens will be sent to the Polio and Picornavirus Laboratory Branch of the Division 
of Viral Diseases, CDC, Atlanta for testing. Sera will be shipped to Atlanta after the completion of the 

 
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18 weeks visits. Stool samples will be shipped to Atlanta in multiple batches. CDC laboratory staff will 
be blinded to the randomization assignment.  
 
Presence of poliovirus neutralizing antibodies to all three poliovirus types will be assessed using 
microneutralization assay (Appendix B). Antibody titers below 1:8 will be considered negative for 
presence of protective levels of poliovirus antibodies. The highest detectable titer will be >1:1448. The 
antibody titers at six weeks of age will be the assumed starting point for the expected decline in 
maternal antibody assuming a half-life of 28 days.  
 
Real-time reverse-transcriptase polymerase chain reaction will be used to detect the presence of type-
specific poliovirus shed in stool samples. For a subset of stool samples, viral recombinants will be 
characterized using high resolution Illumina sequencing (Appendix C). Sequencing reads will be 
mapped against all three Sabin vaccine reference sequences to generate coverage plots.  
 
The figure below provides an overview of specimens that will be collected and tested as part of this 
trial.  
 
 
 
 
Long-term Specimen Storage 
Vaccine recipients will be requested to permit storage of blood and stool specimens for no more than 
five years for potential future use in studies. Prior to testing of the stored specimens for any additional 
objective(s) not specified in the approved protocol, human subject approval would be requested from 
the ethical review committees of icddr,b. Parents of study vaccine recipients will not be contacted, and 
no genetic testing will be performed on blood specimens. All specimens will be destroyed after five 
years from the end date of the study (the date of last clinic visit among study vaccine recipients). 
Specimens will be destroyed per the laboratory’s standard procedures for destruction of biohazardous 
waste.   
 
Sample Size Calculation and Outcome Variables 
Sample size 
Sample size calculations are based on an assumption of 10% attrition and the study is powered to 
address the two primary objectives.  
 

 
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For primary objectives 1 and 2, immune responses to two doses of co-administered nOPV2 and bOPV 
will be assessed for non-inferiority in comparison with two doses of nOPV2 or bOPV given alone. 
Based on unpublished phase 2 data from Panama, we assume that after two doses of nOPV2, 90% of 
vaccine recipients will demonstrate a type 2 immune response. After two doses of bOPV, approximately 
85% of vaccine recipients are expected to demonstrate a type 3 immune response and at least 85% are 
expected to demonstrate a type 1 immune response [24, 25].  
 
A sample size of 238 per group is required to achieve 90% power with α of 0.05 to detect a non-
inferiority margin of 10% in seroconversion proportions between the groups of nOPV2 to nOPV2 with 
bOPV, and bOPV to bOPV with nOPV2. After adjustment for attrition, the enrolment target is 265 per 
group.  
 
Note: For a non-inferiority comparison with a 10% margin, the minimum sample size required for type-
2 response of assumed 90% seroconversion is smaller than that for type-1 and type-3 response of 
assumed 85% seroconversion. As this is a balanced trial, minimum sample size is determined by the 
larger sample size requirement.   
 
To assess transmissibility of nOPV2 among infants receiving one dose of nOPV2 vaccine and their 
siblings, two power calculations were computed to: 1) detect at least one transmission event and 2) 
estimate transmission rate. Both power calculations were based on the established sample size for the 
primary objectives and expected shedding rate. Shedding rates were estimated using mOPV2 shedding 
rates from the mOPV2 transmission study in Bangladesh [26] as a comparator and using a simulation 
model which includes shedding duration and decline in type 2 immunity. All power calculations for the 
simulation are based on the expected shedding at day 14 post-vaccination. As there are two additional 
follow-up points and opportunities to observe transmission, the reported power calculations are likely a 
lower-bound.  
 
Based on the expected recruitment size of 265 vaccine recipients in the nOPV-2 only Arm (Arm A) and 
265 next youngest siblings, many of which are expected to be under the age of 5 years, there is evidence 
of sufficient power to detect transmission events, as well as to estimate a reasonable confidence interval 
for shedding prevalence. Using sample sizes for the primary objectives, the probability of detecting at 
least one household transmission event is close to 1. Using our nOPV2 simulation model, we estimate 
prevalence and 95% CI of shedding to be 53% among vaccine recipients (46.4 – 59.5%) and 23% (17.8 
– 28.8%) among siblings under the age of 5 years.  
 
Table 4: Sample size of study arms 
Arms 
Unadjusted 
sample size 
Enrolment target* 
A 
nOPV2 
238 
265 
B 
nOPV2/bOPV 
238 
265 
C 
bOPV 
238 
265 
Total 
714 
795 
* Adjusted for 10% attrition    
 
Outcome variables 
Poliovirus antibody titers to types 1, 2 and 3 will be measured in sera extracted from blood collected at 
6, 10, 14, and 18 weeks of age. Seropositivity is defined as reciprocal polio neutralizing antibody titers 
of at least 1:8. Immune response will be defined as either seronegative vaccine recipients (<1:8 titers) 

 
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who become seropositive (≥1:8) (seroconversion) or vaccine recipients who demonstrate at least a four-
fold increase in titer (boosting). Baseline maternal antibody titer will be determined at 6 weeks of age 
and the estimated maternal antibody level at each blood collection will be calculated assuming an 
exponential decline with a half-life of 28 days. Vaccine recipients and siblings will be categorized for 
shedding poliovirus through results from multiple real-time PCR (rRT-PCR) assay designed to detect 
poliovirus. Household transmission will be defined as present if siblings test positive for nOPV2 
vaccine virus at either 1, 2, or 4 weeks following the vaccine recipient’s vaccination at 6 weeks of age. 
nOPV2 vaccine virus in stool specimens from vaccine recipients and siblings will be further 
characterized through high-resolution Illumina RNA sequencing. Intertypic vaccine-derived 
recombinants will be characterized in stool collected from study vaccine recipients and siblings. 
 
Randomization 
Vaccine recipients will be randomized to one of three arms using a block randomization scheme. The 
randomization algorithm will be generated by CDC staff and executed in REDCap. The randomization 
algorithm will not be known to study staff in icddr,b. Randomization assignment will be revealed after 
completing enrolment procedures, when study staff “click” on randomization button on the study 
tablets. Therefore, study staff enrolling and managing vaccine recipients will not have a priori 
knowledge of randomization scheme. As this is an open-label trial, the arm assignment of vaccine 
recipients will be known to study staff and the vaccine recipient’s parents after randomization. 
Laboratory staff processing blood samples in Bangladesh will not have a priori knowledge of 
randomization scheme but will know the arm assignment after randomization. However, CDC 
laboratory staff testing for poliovirus antibody titers will remain blinded throughout the study.  
 
Timeline of activities 
Activities 
Year 1 (months) 
2 
4 
6 
8 
10 
12 
Protocol development/IRB approval, 
Staff training/site preparation 
 
 
 
 
 
 
Enrolment 
 
 
 
 
 
 
Start and completion of vaccination 
 
 
 
 
 
 
Follow up of vaccine recipients 
 
 
 
 
 
 
Lab testing  
 
 
 
 
 
 
Dissemination/Publications 
 
 
 
 
 
 
 
Data Analysis 
General Issues 
This is an open-label, non-inferiority, randomized clinical trial that will compare immune response to 
types 1, 2 and 3 polioviruses of nOPV2 and bOPV when co-administered compared to nOPV2 and 
bOPV given alone. The study will recruit 795 vaccine recipients in three arms. Vaccine recipients will 
be enrolled at 6 weeks of age and followed until 18 weeks of age, for 12 weeks of follow-up. For each 
vaccine recipient enrolled, one sibling will be enrolled at the same time and followed for 4 weeks .  
 
General Analytical Issues 
The data will be analysed by study investigators that include epidemiologists and statisticians. 
Statistical packages that will be used for the analysis include R, SAS and STATA. 
 
For assessing immunity to polio, seropositivity is defined as reciprocal polio neutralizing antibody titers 
 
 of at least 1:8. Immune response will be defined as either seronegative vaccine recipients (<1:8 titers) 
who become seropositive (≥1:8) [seroconversion] or vaccine recipients who demonstrate a four-fold 

 
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increase in titer (boosting). Baseline maternal antibody titers will be determined at 6 weeks of age and 
the estimated maternal antibody level at each blood collection will be calculated assuming an 
exponential decline with a half-life of 28 days.  
 
For assessing fecal viral shedding, detection of nOPV2 or bOPV vaccine virus in stool specimens from 
vaccine recipient or sibling at either 1, 2, or 4 weeks post-vaccination of the vaccine recipient at 6 
weeks of age will be defined as positive for viral shedding. For assessing the detection of household 
transmission, presence of transmission will be defined as the sibling of the vaccine recipient testing 
positive for nOPV2 vaccine virus at either 1, 2, or 4 weeks post-vaccination of the vaccine recipient at 6 
weeks of age. 
 
To characterize the intertypic recombinants in a subset of stool specimens of vaccine recipients and 
siblings after one dose of nOPV2 given with or without bOPV, amplicon sequences from long-read 
sequencing methods will be used to confirm genome structure. Amplicon sequences will also be 
mapped against all three Sabin vaccine reference sequences to generate coverage plots. 
 
Primary Analytical Approach 
The primary analytical approach will be intention-to-treat. To evaluate the primary objectives, the 
intention-to-treat analysis will be restricted to vaccine recipients who: 
• Have not withdrawn consent for receipt of vaccines and follow-up evaluations since enrolment. 
• Have adequate blood specimen for serological analysis from the enrolment visit at 6 weeks of 
age. 
• Have adequate blood specimen for serological analysis from the follow-up visit as mentioned by 
arm by primary objective in Table 5. 
 
For secondary objectives 1 and 2, the intention-to-treat analysis will be restricted to vaccine recipients 
who: 
• Have not withdrawn consent for receipt of vaccines and follow-up evaluations since enrolment. 
• Have adequate blood specimen for serological analysis from the enrolment visit. 
• Have adequate blood specimen for serological analysis from the follow-up visit as mentioned by 
arm by secondary objective in Table 5. 
 
For secondary objectives 3-5, the intention-to-treat analysis will be restricted to vaccine recipients and 
siblings who: 
• Have not withdrawn consent for receipt of vaccines and follow-up evaluations since enrolment. 
• Have adequate stool specimens to examine vaccine-virus particles. 
 
Secondary Analytical Approach 
In addition to an intention-to-treat analysis, we will also perform per protocol analysis. To evaluate the 
primary objectives, per protocol analysis will be limited to vaccine recipients with the same restrictions 
as that outlined in primary analytical approach for primary objectives with additional restrictions of: 
• Adequate blood specimen for serological analysis within 3 days of the scheduled visit date. This 
applies to enrolment visit blood specimen as well as follow-up visit blood collection. 
• Received all scheduled study vaccines within 3 days of scheduled visit date. 
 
For secondary objectives 1 and 2, the per protocol analysis will be limited to vaccine recipients with the 
same restrictions as those outlined in the primary analytical approach for secondary objectives 1 and 2 
with additional restrictions of: 

 
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• Adequate blood specimen for serological analysis within 3 days of the scheduled visit date. This 
applies to enrolment visit blood specimen as well as follow-up visit blood collection.  
• Received all scheduled study vaccines within 3 days of scheduled visit date. 
 
For secondary objectives 3-5, the per protocol analysis will be limited to vaccine recipients or siblings 
with the same restrictions as those outlined in the primary analytical approach for secondary objectives 
3-5 with additional restrictions of: 
• Adequate stool specimen to examine vaccine-virus particles within 3 days of the scheduled visit 
date.  
• Vaccine recipient received the scheduled study vaccination at 6 weeks of age. 
` 
Table 5: Study arms comparison, study endpoints, and results by objective 
 
Study Objective 
Study Arms 
Comparison 
Endpoint (vaccine 
recipient age) 
Results Included in 
Analysis 
Primary 
  
  
  
1.  To compare type 2 immunogenicity of 
two doses of nOPV2 with two doses of 
nOPV2 co-administered with bOPV. 
A vs. B 
14 weeks 
Serology: 6, 10, 14 weeks 
2.  To compare type 1 and 3 immunogenicity 
of two doses of bOPV with two doses of 
bOPV co-administered with nOPV2. 
C vs. B 
14 weeks 
Serology: 6, 10, 14 weeks 
Secondary 
  
  
  
1.  To compare type 2 immunogenicity of 
one or three doses of nOPV2 with the same 
number of doses of nOPV2 co-administered 
with bOPV. 
A vs. B 
10 weeks 
Serology: 6, 10 weeks 
18 weeks 
Serology: 6, 10, 14, 18 
weeks 
2.  To compare type 1 and 3 immunogenicity 
of one or three doses of bOPV with the same 
number of doses of bOPV co-administered 
with nOPV2. 
C vs. B 
10 weeks 
Serology: 6, 10 weeks 
18 weeks 
Serology: 6, 10, 14, 18 
weeks 
3. To assess the presence of fecal viral 
shedding (as determined by real-time RT-
PCR) over time following one dose of 
nOPV2, bOPV, or co-administered nOPV2 
and bOPV.   
Descriptive 
with 
comparisons 
of A vs. B 
and B vs. C 
10 weeks (i.e. 4 
weeks post-
vaccination) 
 
 
Stool: 7 (Arm A only), 8, 10 
weeks from vaccine recipient  
4. To estimate household transmission of 
nOPV2 by assessing the presence of fecal 
viral shedding (as determined by real-time 
RT-PCR) among siblings of vaccine 
recipients.  
Descriptive  
10 weeks (i.e. 4 
weeks post-
vaccination) 
 
Stool: 7 (Arm A only), 8, 10 
weeks from sibling 
5. To assess the occurrence and genomic 
characteristics of intertypic recombinants in 
stool specimens of vaccine recipients and 
siblings after one dose of nOPV2 given with 
or without bOPV. 
Descriptive 
with 
comparison 
of A vs. B 
10 weeks 
(i.e. 4 weeks post-
vaccination) 
Stool: 8, 10 weeks from 
vaccine recipient and sibling 
 
 
Adverse Events (AEs) 
Safety will be assessed by: 

 
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• Incidence rate of solicited systemic AEs 24–48 hours after the first study vaccination. 
• Incidence rate of unsolicited AEs during entire study period.  
• Incidence rate of serious AEs during entire study period. 
 
Data Safety Monitoring Plan (DSMP) 
Records to be kept 
The study will use a mix of paper- and electronic-based records. Participant data from clinic visits (for 
vaccine recipients and from siblings, who will attend the first clinic visit only) will be collected and 
recorded electronically on tablets through REDCap, an electronic data entry system. The Informed 
Consent Form and Screening Form will be paper-based exclusively. Eligibility Criteria, which is 
assessed at the beginning of study form Clinic Visit at 6 weeks will be exclusively paper-based for 
participants who are not eligible or those who are eligible but did not consent. For those who are 
eligible and have provided consent for participating in the study, the Eligibility Criteria component of 
study form Clinic Visit at 6 weeks will be paper- and web- based. Female field workers will use paper-
based forms for the home visits; these will be entered by data entry staff into REDCap. All other study 
information for participants will be exclusively web-based. As a back-up, there will be paper-based 
CRFs available to collect participant data in case of any problems that arise with the web-based system. 
Paper-based data collection will be used as back-up in the case of power failure or malfunctioning WiFi 
or tablets during study procedures. In case there is complete failure of tablet-based system, the study 
will switch to paper-based CRF.   
 
Vaccine recipients will be identified by a PID, which will be provided after enrolment and 
randomization. The PID will be used to link information recorded in eCRFs and results from laboratory 
analysis of biological specimens. Personal identifiers will only be recorded on the Screening Form and 
the Informed Consent Form but will not be recorded on any eCRFs, biological specimens, or SAE 
forms. The two paper-based documents (Screening Form and Informed Consent Form) will be filed in a 
locked cabinet separately from the other study forms and logs. Information about the sibling and his/her 
stool specimens will be captured under the vaccine recipient’s PID.  
 
Data Entry, Cleaning and Storage 
Paper records will be kept in a locked cabinet, in a locked room, with access restricted to select study 
staff and investigators. The computers and tablets will be kept in a locked room and password protected. 
Only select study staff and investigators will be authorized to access the digital records. Laptops/tablets 
will be password protected with two-levels of password protection. The first password will be device 
password. The second password will be to log in to the web-based data entry portal.  
 
Information collected in the CRFs other than Screening Form will be entered into a web-based data 
collection system (REDCap). The web-based data collection system will include system data checks to 
identify and prevent data entry errors. Data collected from the web-based data collection tools will be 
stored in secure server accessible only after providing user ID and password. After the completion of 
analysis and publication of study results, the study forms and databases will be archived. The forms will 
be stored in a locked cabinet with access restricted to investigators. The database will be password 
protected. 
 
Data Ownership and Public Release 
Study data will be jointly owned by icddr,b and CDC. After completion of study activities, summary 
tables of study results will be available on www.clinicaltrials.gov. After study registration, 
clinicaltrials.gov provides unrestricted public access on study details including summary tables. 
 

 
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Study investigators will have access to all participant (vaccine recipient and sibling) data, without 
identifiers, including laboratory data. Co-investigators and technical advisors for the household 
transmission objectives will have access to relevant data as defined in Table 5 for the corresponding 
objectives.  
 
Site Monitoring Plan 
Prior to start of the study, Standard Operating Procedures (SOPs) will be developed by investigators in 
consultation with study staff. Study staff will receive training on the SOPs for study procedures and on 
Good Clinical Practices in clinical trials.  
After the start of the study, monitoring will be conducted four times: 
1. First monitoring visit: within 2 months after the start of the study 
2. Second monitoring visit: within 2 months after the first monitoring visit 
3. Third monitoring visit: within 2 months after the second monitoring visit 
4. Final monitoring visit: Within two months of the last study visit by vaccine recipients  
 
During these monitoring visits, the following study documents and procedures will be reviewed: 
1. Participant records, including informed consent forms (with assistance of a medical officer) 
2. eCRFs, electronic records, and other supporting data 
3. Adverse event forms (with assistance of a medical officer) 
4. DSMB reports 
5. Laboratory specimen records 
6. Vaccine and biological materials storage and records 
7. Any additional medical records (with assistance of a medical officer) 
8. Measures to ensure protection of study participants 
9. Measures to ensure compliance with study protocol, and accuracy and completeness of records 
10. Any regulatory files associated with the study will also be inspected to ensure all regulatory and 
reporting requirements are being followed 
 
Site monitoring visits will be conducted by CDC staff who have not interacted with study participants. 
Site monitoring reports will be shared with the PI.   
 
Safety Monitoring 
CDC and icddr,b have conducted several recent polio clinical trials using OPV and IPV (i.e., tOPV, 
bOPV, mOPV2, fIPV, and IPV) that have included safety monitoring of participants.   
  
Anaphylactic reactions to bOPV are very rare but possible because bOPV contains trace amounts of 
antibiotics, including kanamycin and erythromycin for bOPV manufactured by Bio Farma. The nOPV2 
being used in this trial does not contain antibiotics. All vaccine recipients will be observed for 30 minutes 
after vaccination to monitor for any immediate adverse reactions to the vaccine. Properly skilled medical 
personnel will be immediately available in the event of an unexpected adverse reaction. Adrenaline (for 
anaphylactic reaction) will be available but equipment for endotracheal intubation will not be available at 
the site. After the 30-minute observation period, the medical officer will record the presence of any 
potential reaction to the vaccine on the corresponding CRF. At the end of each study visit, the physician 
will provide the parents with a phone number to call if they have any questions or if the vaccine recipient 
experiences any reactions to the vaccines. Since nOPV2 is a new vaccine, as an additional safety 
monitoring measure, study staff will contact the household of vaccine recipients to inquire and record any 
potential adverse events after the first vaccination visit; this will take place 24–48 hours after vaccination.  
 
Should a serious illness occur while the vaccine recipient is enrolled in the study (requiring a physician's 
visit or hospitalization), parents will be instructed to seek medical care immediately and to notify study 

 
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staff as soon as possible. Medical care for vaccine recipients will be provided free of charge for expected 
minor illnesses that develop during the follow up period, such as diarrhea and respiratory infections, as 
well as any adverse outcomes judged to be possibly, probably or definitely related (detailed below) to 
study vaccines or vaccination.  
 
Recording, Monitoring and Reporting of Adverse Events 
 
Solicited AEs: In this study, fever, vomiting, crying abnormal, drowsiness, loss of appetite and 
irritability will be solicited at one timepoint 24-48 hours after the first study vaccination. This will 
include the definitions of mild, moderate and severe AEs, as described in Table 6, to facilitate the 
assessments of the level of functional impairment for each experienced AE. For fever, temperature will 
be recorded if available, otherwise subjective fever will be solicited.  
 
Table 6:  Intensity scales for solicited symptoms 
Events 
Severity Grade 
0 
1 
2 
3 
Fever (if 
thermometer is 
available) 
<37.5° C 
37.5° C- 38.0 ° C 
38.1° C- 39.0 ° C 
>39.0 ° C 
Vomiting 
None 
1 episode/24 hours 
2-5 episodes/24 
hours 
≥ 6 episodes/24 
hours 
Abnormal 
Crying 
None 
< 1hour 
1-3 hours 
>3 hours 
Drowsiness 
None 
Sleepier than usual 
or less interested in 
surroundings 
Not interested in 
surroundings or did 
not wake up for a 
feed 
Sleeping most of the 
time or difficult to 
wake up 
Poor feeding 
None 
Eating less than 
normal 
  
Missed 1 or 2 feeds 
completely 
Refuses ≥3 feeds or 
refuses most feeds 
Irritability 
None 
  
Easily consolable 
  
Requiring 
increased attention 
  
Inconsolable 
 
Unsolicited AEs: The investigator will assess the incidence and maximum intensity that occurred over 
the duration of the event for all unsolicited AEs recorded during the study. The assessment will be 
based on the investigator’s clinical judgment. The intensity should be assigned to one of the following 
categories: 
1 (mild) = An AE which is easily tolerated by the subject, causing minimal discomfort and not 
interfering with everyday activities. 
2 (moderate) = An AE which is sufficiently discomforting to interfere with normal everyday 
activities. 
3 (severe) = An AE which prevents normal, everyday activities.  
 
During each study clinic visit, study staff will question the parents about possible AEs experienced by the 
vaccine recipient since the previous study visit. This information will be recorded on the corresponding 

 
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eCRF. Reported AEs will be recorded in the AE form in the eCRF. The AE form will include a description 
of the event, time of onset, assessment of severity, relationship to study product, and time of 
resolution/stabilization of the event.  
  
Reports of AEs will be periodically discussed by the team of investigators. The study team will review 
the potential relationship and classify the relationship into:  
-        Unrelated 
-        Possibly related 
-        Probably related 
-        Definitely related 
  
Interpretation of vaccine-relationship to AE will be based on the type of event, the relationship of the 
event to the time of vaccine administration, the known biology of the vaccine and the investigators’ 
medical judgment. In addition, the investigation team will also report clusters of AE (at least three) to the 
DSMB periodically for further evaluation. 
  
An AE will be considered a serious AE (SAE) if it meets any of the following criteria: 
1. Death during the study period 
2. Life threatening event 
3. Hospitalization or prolongation of existing hospitalization 
4. Paralysis, severe disability/incapacity, or substantial disruption of the ability to conduct 
normal functions 
5. Anaphylaxis associated with vaccine administration  
  
All SAEs will be notified by the principal investigator to the regulatory agencies, ethical review 
committees, the Data Safety Monitoring Board (DSMB), and CDC within 24 hours of information. A 
separate icddr,b SAE form will be completed for SAEs and submitted to the ERC for review. 
 
Data Safety Monitoring Board (DSMB) 
The study will be monitored by a DSMB constituted by icddr,b with input from the Ethical Review 
Committee (ERC). The DSMB will include representation outside of icddr,b. The DSMB is expected to 
convene once prior to the start of the study after ethical and regulatory approval of the study protocol. 
The DSMB will convene meetings at the middle and at study completion. The DSMB will be 
responsible for establishing study stopping rules. Immunogenicity data will not be available for DSMB 
meetings because all blood specimens will be analysed after the completion of the field activities. 
 
 
 
Ethical Assurance for Protection of Human rights 
Justification for Involving a Vulnerable Population 
Vaccine recipients will be enrolled at 6 weeks of age. Enrolment at this early age is required because the 
study is assessing the immunogenicity of poliovirus vaccines that are routinely administered at this age. 
The current schedule for polio immunization in Bangladesh includes three doses of bOPV administered 
at 6, 10 and 14 weeks of age plus two fractional IPV (fIPV) ID doses administered at 6 and 14 weeks of 
age. Delaying vaccine recipient enrolment until they are older will not allow assessing the 
immunogenicity of alternative polio vaccines and vaccination schedules because most vaccine 
recipients will be immune as a result of the polio vaccines they received through routine immunization. 
 
Potential Benefits and Compensation to Vaccine recipients 
Vaccine recipients will receive all non-polio routine immunizations (BCG, Penta, PCV) as per the routine 
immunization schedule of Bangladesh. After completion of study activities, vaccine recipients in Arms 
A and B will receive three doses of bOPV. The three doses of bOPV will be given in 4-week intervals 

 
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beginning at 18 weeks of age. All vaccine recipients will also receive two doses of fIPV. The routine 
polio immunization schedule in Bangladesh is three doses of bOPV along with two doses of fIPV. During 
study visits, vaccine recipients will be examined by medical officers and their growth and development 
will be assessed. In addition, vaccine recipients and siblings will be offered advice on diagnosis and 
treatment of any other illnesses they experience during the study period at no cost. 
  
For the larger public health community, the results of this clinical trial may be used to inform outbreak 
response vaccination strategies. 
 
Risks to Vaccine recipients and Siblings 
bOPV has a long safety record and is used for routine childhood immunization in Bangladesh; nOPV2 is 
similar to mOPV2 except that it has further attenuated type 2 poliovirus. mOPV2 has been used in a polio 
vaccine trial by icddr,b in 2015–2016. Compared to mOPV2 and other existing OPVs, nOPV2 has been 
developed to be more genetically stable and therefore less likely to revert to neurovirulence. Phase 1 
clinical trials of nOPV2 in IPV-primed adults demonstrated the safety of both vaccine candidates [14]. 
For both, viral shedding appeared to be no greater than mOPV2, and possibly lower. Mouse 
neurovirulence testing of shed virus also demonstrated the stability of the shed virus and deep sequencing 
of the shed virus revealed no unexpected mutations or loss of key stabilizing mutations, which implies a 
decreased likelihood of transmission of a virus with neurovirulent phenotype, relative to Sabin 2. 
Preliminary results from phase 2 trials in Belgium and Panama were reviewed by an independent data 
safety monitoring board, which supported age group progression from adults to toddlers and infants based 
on the available safety information. Early trial results also suggest that both candidates are immunogenic 
and more genetically stable than Sabin 2 [16]. Prior to the start of the current study, icddr,b will implement 
an OPV-naïve infants trial in Matlab that includes nOPV2 (PR-20001).   
 
Vaccine recipients may have less immunity to polio during the study period than similarly aged non-
vaccine recipients because of the altered vaccination schedule; all vaccine recipients will receive their 
first dose of fIPV twelve weeks later than the EPI schedule and some vaccine recipients will receive 
their first dose of bOPV twelve weeks later than the EPI schedule. However, the risk of infection with 
WPV or cVDPV is very low in Bangladesh. The last endemic WPV case was reported in 2000, and the 
last imported case was reported in 2006. Furthermore, WPV type 2 and WPV type 3 have been declared 
globally eradicated. No cVDPV cases have ever been reported in Bangladesh thanks to the high 
coverage reached with routine immunization since WPV elimination. 
 
The most significant adverse event associated with OPV is vaccine-associated paralytic poliomyelitis 
(VAPP); however, the risk for recipient VAPP and contact VAPP is very small as the children will be 
protected by maternal antibodies. The risk of VAPP is about 1 case per 1.2 to 2.4 million doses of tOPV 
[3]; the risk after bOPV is expected to be similar or slightly higher than tOPV for recipient VAPP as 
most were related to types 1 or 3 [4-6]. Because bOPV is given as part of the EPI schedule in 
Bangladesh, study vaccine recipients will have similar risk for developing VAPP to types 1 and 3 as 
infants not participating in the study. There may also be some risk of VAPP in close contacts of vaccine 
recipients, including the older siblings, but we assume this risk to be minimal in Bangladesh as 
Bangladesh has high routine immunization coverage and nOPV2 has been designed with the intent to be 
much less prone to reversion to neurovirulence compared to Sabin OPV.   
 
Upon completion of study activities, vaccine recipients in the nOPV2-only arm will not have protection 
against types 1 and 3 and vaccine recipients in the bOPV-only arm will not have protection against type 
2 until they receive their first dose of fIPV (and bOPV, depending on the arm) at 18 weeks of age. 
 

 
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In summary, because of protection conferred by the nOPV2 and bOPV doses provided by the study, the 
genetic stability of nOPV2, and the low probability of poliovirus circulation in Bangladesh during the 
study implementation, the risk of paralysis caused by WPV, cVDPV or OPV in study vaccine 
recipients, will be very low and similar or only slightly different than the risk of non-vaccine recipient 
infants of similar age. 
 
Additional potential risks and discomfort to vaccine recipients stem from blood collections, adverse 
reactions to study vaccines, and breach of confidentiality or an unintended disclosure of confidential 
information. Immunogenicity to study vaccines can only be assessed from blood specimens. Four blood 
draws are necessary because immunogenicity is measured by determining the change in antibody titers. 
The subsequent blood draws 4 weeks after administration of study vaccines will assess immunogenicity 
of study vaccines per the study objectives. The minimal amount of blood necessary for testing (1 ml) 
will be collected with the most common side effect being pain at the site of collection. To minimize the 
discomfort and risks associated with blood draws, only qualified staff with experience in the collection 
of blood from infants will draw blood. Sterile equipment and technique, and alcohol swabs will be used 
to reduce the risk of infection from a blood draw. Pressure will be applied at the site of blood draw to 
minimize bleeding from the site of blood draw. 
 
bOPV contains trace amounts of antibiotics; therefore, vaccine recipients may react to these substances 
although reactions are rare. To minimize the consequences of a potential anaphylactic reaction to study 
vaccines and/or their components, vaccine recipients will be observed for 30 minutes after vaccine 
administration by staff trained to respond to potential reactions. Parents of the vaccine recipient will 
also be contacted (either via phone call or home visit) 24–48 hours after the first vaccination. The 
purpose will be to inquire about and record any potential adverse events since nOPV2 is a new vaccine.  
 
An additional risk to participants is a breach of confidentiality or an unintended disclosure of personal 
information. All study personnel will be trained in the protection of human subjects in research. All 
Case Report Forms (CRFs), laboratory specimens, and other reports including adverse events will be 
identified by a Participant Identification Number (PID) to maintain participant confidentiality. No 
identifiers will be recorded on eCRFs, other study reports and laboratory specimens. All paper records 
will be stored in a locked cabinet. All electronic records will be stored in a password protected database, 
on a password protected computer. Access to paper and electronic records will be restricted to study 
staff and investigators. De-identified data may be shared with vaccine manufacturers and regulatory 
authorities, both national and international, upon request. 
 
Institutional Review Board (IRB) and Ethical Review 
The protocol with letters of consent and other associated appendices will be reviewed and approved by 
the Research Review Committee (RRC) and Ethical Review Committee (ERC) and at icddr,b before 
any participants are enrolled. The RRC and ERC at icddr,b will be responsible for oversight of the 
study. Any subsequent modifications to the protocol will be submitted to the ERC and RRC at icddr,b 
for additional approval. 
 
All participants will be below the legal age of consent because vaccine recipient participants will be 
enrolled at 6 weeks of age and siblings will be <10 years of age. Parents of potential participants will be 
informed that their children’s participation in the study is strictly voluntary and that they are free to 
withdraw their children from the study at any time. Signed informed consent will be obtained from a 
parent of the participants, in the presence of a literate witness who is not directly related to the study. 
The informed consent form will describe the purpose of the study, procedures, confidentiality of 
information, and the risks and benefits of participation. Informed consent forms will be translated from 
English to Bangla, the local language. A copy of the informed consent form will be given to the parent 

 
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of the participants and noted in study records. A second copy of the signed informed consent form will 
be filed with study records.  
 
Participant Confidentiality Procedures 
All eCRFs, laboratory specimens, and other reports including adverse events will be identified by PID 
to maintain participant confidentiality. No identifiers will be recorded on eCRFs, other study reports 
and laboratory specimens. All paper records will be stored in a locked cabinet. All electronic records 
will be stored in a password protected database, on a password protected computer. Access to paper and 
electronic records will be restricted to study staff and investigators; CDC co-investigators will only 
have access to de-identified electronic records. De-identified data may be shared with vaccine 
manufacturers and regulatory authorities, both national and international, upon request.  
 
Use of Animals 
Not applicable 
 
Collaborative Arrangements 
This study includes two partners with the following roles in the study:  
1. International Centre for Diarrhoeal Diseases Research, Bangladesh 
• Design of the study and collaborate in the development of the study protocol  
• Submission of protocol to icddr,b’s Research and Ethical Review Committees.  
• Select study site and study staff and develop a budget for study implementation. 
• Work with vaccine manufacturers to secure material transfer agreements for study vaccines, if 
needed. 
• Collaborate in the development of standard operating procedures, study forms and training material.  
• Conduct training and supervise study staff. 
• Carry out implementation of the study procedures and coordinate logistics for sample collection, 
processing, storage and shipment as well as study vaccine storage and administration.  
• Record and provide treatment for potential adverse events developed during the study. 
• Create and enter data from case report forms, if paper-based forms used for any participants, into a 
database for future analysis.  
• Collaborate with CDC investigators in the analysis, interpretation and writing of study results for 
study reports and manuscripts to be published in peer reviewed journals. 
 
2. Centers for Disease Control and Prevention, Atlanta, GA, USA  
• Design of the study and lead in the development of the study protocol.  
• Submission of the protocol for CDC ethical clearance.  
• Provision of funding for study implementation.    
• Provide support to icddr,b to obtain material transfer agreements for study vaccines from vaccine 
manufacturers, if needed. 
• Collaborate in the development of standard operating procedures, study forms and training material.  
• Provide support in the training of study staff. 
• Plan vaccine recipient randomization assignment for vaccine recipient assignment during enrolment.  
• Monitor study implementation through visits to the study site in coordination with icddr,b. 
• Test serum specimens for the determination of antibodies against poliovirus and enter laboratory 
data into an electronic database. 
• Test stool specimens for the presence of vaccine polioviruses and recombinants.  
• Lead data analysis, interpretation of results and writing detailed reports and manuscripts for 
publication in peer reviewed journals in collaboration with icddr,b investigators. 
 

 
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Facilities Available 
Clinical Facilities 
The International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b) has a large multi-
disciplinary international and national scientific research staff. Existing field, hospital, laboratory and 
office facilities will be used for this study. icddr,b scientists have conducted a variety of vaccine studies 
including poliovirus. 
 
Field Site 
The study will be carried out in urban slums in Mirpur in Dhaka. CDC and icddr,b have previously 
collaborated on seven polio clinical trials at Mirpur.  
 
If feasible based on guidelines/recommendations related to the COVID-19 pandemic (e.g., physical 
distancing), the study will also be conducted at CTU, with the adjoining slum and non-slum areas of 
icddr,b Mohakahli, Dhaka - Kunipra, Arjat Para, Nakhalpara, Korail, Begunbari, Chairman bari and 
Badda. CDC and icddr,b have previously collaborated on two polio clinical trials at Mohakahli.  
 
Laboratory Facilities at icddr,b 
Existing laboratory facilities in icddr,b will be used to store and process specimens prior to shipment to 
CDC, Atlanta. 
 
Laboratory Facilities at CDC 
The Polio and Picornavirus Laboratory Branch of the Division of Viral Diseases at CDC, Atlanta has the 
necessary staff, infrastructure and equipment to perform the laboratory testing for this study. The testing 
of serum and stool using the methods outlined in this protocol requires specific training and equipment 
that is not currently available at the laboratory of the Institute of Epidemiology, Disease Control and 
Research or at any other laboratory in Bangladesh.   
 
 
Literature Cited 
 
[1] Nathanson N, Kew OM. Poliovirus vaccines: past, present, and future. Arch Pediatr Adolesc Med. 
2011;165:489-91. 
[2] Sutter R, Kew O, Cochi S, Aylward RB. Poliovirus vaccine -- live. In: Plotkin S, Orenstein WA, Offit 
PA, editors. Vaccines. 6 ed. Philadelphia: Elsevier Saunders; 2012. 
[3] Dowdle WR, De Gourville E, Kew OM, Pallansch MA, Wood DJ. Polio eradication: the OPV 
paradox. Rev Med Virol. 2003;13:277-91. 
[4] Estivariz CF, Pallansch MA, Anand A, Wassilak SG, Sutter RW, Wenger JD, et al. Poliovirus 
vaccination options for achieving eradication and securing the endgame. Current opinion in virology. 
2013;3:309-15. 
[5] Burns CC, Diop OM, Sutter RW, Kew OM. Vaccine-derived polioviruses. J Infect Dis. 2014;210 
Suppl 1:S283-93. 
[6] The Global Polio Eradication Initiative. Polio Eradication and Endgame Strategic Plan 2013-2018. 
Geneva, Switzerland2013. 
[7] Initative TGPE. Polio Eradication and Endgame Strategic Plan 2013 - 2018. Geneva, Switerland2013. 
[8] Vidor E, Plotkin S. Poliovirus vaccine - inactivated. In: Plotkin S, Orenstein WA, Offit PA, editors. 
Vaccines. 6 ed. Philadelphia: Elsevier Saunders; 2012. 
[9] Centers for Disease Control Prevention. Apparent global interruption of wild poliovirus type 2 
transmission. MMWR Morbidity and mortality weekly report. 2001;50:222-4. 
[10] Adams A, Salisbury DM. Eradicating polio. Science. 2015;350:609. 

 
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[11] World Health Organization. Meeting of the Strategic Advisory Group of Experts on Immunization, 
April 2013 -  Conclusions and Recommendations. Wkly Epidemiol Rec. 2013;88:201-16. 
[12] Te Yeh M, Bujaki E, Dolan PT, Smith M, Wahid R, Konz J, et al. Engineering the live-attenuated 
polio vaccine to prevent reversion to virulence. 2020. 
[13] Konopka-Anstadt JL, Campagnoli R, Vincent A, Shaw J, Wei L, Wynn NT, et al. Development of a 
new oral poliovirus vaccine for the eradication end game using codon deoptimization. 2020;5:1-9. 
[14] Van Damme P, De Coster I, Bandyopadhyay AS, Revets H, Withanage K, De Smedt P, et al. The 
safety and immunogenicity of two novel live attenuated monovalent (serotype 2) oral poliovirus vaccines 
in healthy adults: a double-blind, single-centre phase 1 study. 2019. 
[15] Belgian Biosafety Server. https://www.biosafety.be/content/phase-2-vaccine-study-healthy-adults-
and-adolescents-evaluate-safety-and-immunogenicity-two. Accessed January 20, 2020. 
[16] Bandyopadhyay A. SAGE Meeting, October 2019: Polio the last mile-Session 9. 
https://www.who.int/immunization/sage/meetings/2019/october/2_nOPV2__SAGE_vf.pdf?ua=1. 
Accessed January 20, 2020. 
[17] Combelas N, Holmblat B, Joffret M-L, Colbère-Garapin F, Delpeyroux FJV. Recombination 
between poliovirus and coxsackie A viruses of species C: a model of viral genetic plasticity and 
emergence. 2011;3:1460-84. 
[18] Cammack N, Phillips A, Dunn G, Patel V, Minor PJV. Intertypic genomic rearrangements of 
poliovirus strains in vaccinees. 1988;167:507-14. 
[19] Lipskaya GY, Muzychenko A, Kutitova OK, Maslova SV, Equestre M, Drozdov S, et al. Frequent 
isolation of intertypic poliovirus recombinants with serotype 2 specificity from vaccine‐associated polio 
cases. 1991;35:290-6. 
[20] Furlone M, Guillot S, Otelea D, Balanant J, Candrea A, Crainic RJV. Polioviruses with natural 
recombinant genomes isolated from vaccine-associated paralytic poliomyelitis. 1993;196:199-208. 
[21] Georgescu M-M, Delpeyroux F, Tardy-Panit M, Balanant J, Combiescu M, Combiescu AA, et al. 
High diversity of poliovirus strains isolated from the central nervous system from patients with vaccine-
associated paralytic poliomyelitis. 1994;68:8089-101. 
[22] Georgescu M-M, Delpeyroux F, Crainic RJJoGV. Tripartite genome organization of a natural type 
2 vaccine/nonvaccine recombinant poliovirus. 1995;76:2343-8. 
[23] Estivariz CF, Anand A, Gary HE, Jr., Rahman M, Islam J, Bari TI, et al. Immunogenicity of three 
doses of bivalent, trivalent, or type 1 monovalent oral poliovirus vaccines with a 2 week interval between 
doses in Bangladesh: an open-label, non-inferiority, randomised, controlled trial. Lancet Infect Dis. 
2015;15:898-904. 
[24] Anand A, Zaman K, Estivariz CF, Yunus M, Gary HE, Weldon WC, et al. Early priming with 
inactivated poliovirus vaccine (IPV) and intradermal fractional dose IPV administered by a microneedle 
device: A randomized controlled trial. Vaccine. 2015;33:6816-22. 
[25] Sutter RW, John TJ, Jain H, Agarkhedkar S, Ramanan PV, Verma H, et al. Immunogenicity of 
bivalent types 1 and 3 oral poliovirus vaccine: a randomised, double-blind, controlled trial. 
2010;376:1682-8. 
[26] Taniuchi M, Famulare M, Zaman K, Uddin MJ, Upfill-Brown AM, Ahmed T, et al. Community 
transmission of type 2 poliovirus after cessation of trivalent oral polio vaccine in Bangladesh: an open-
label cluster-randomised trial and modelling study. 2017;17:1069-79. 
 
 
 

 
46 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Budget 
 
 
Name of the Project/Protocol: 
Immunogenicity of novel-monovalet OPV type 2 (nOPV2) 
Budget Period:
Sept. 30, 2020 to Sept. 29, 2021
Name of the PI:
Dr. K. Zaman
Particulars
Pay 
level
 Month 
Rate (S) 
# of 
Staff
% Time
 # of 
Month 
 Inflation 
@ 8% 
Total Cost 
US$
Personnel: 
Dr. K. Zaman
Intl
15,625
    
1
15%
12
1.08
30,375
         
Dr. Md. Yunus
Intl
13,500
    
1
5%
12
1.08
8,748
           
Study Coordinator/Research Investigator
NOA
2,224
      
1
50%
12
1.08
14,412
         
Field Research Officer
GS5
1,010
      
1
100%
12
1.08
13,090
         
Field Research Supervisor
GS4
788
         
3
100%
12
1.08
30,637
         
Field Research Assistant
GS3
643
         
3
100%
12
1.08
25,000
         
Female Field Worker / SW / FA
UNCL
292
         
18
100%
12
1.08
68,118
         
Medical Officer
NOA
2,056
      
3
100%
12
1.08
79,937
         
Nurse
GS5
1,010
      
3
100%
12
1.08
39,269
         
Senior Scientific Officer
NOA
2,308
      
1
50%
12
1.08
14,956
         
Senior Research Officer (Lab)
GS6
1,306
      
1
100%
12
1.08
16,926
         
Research Officer (Lab)
GS5
1,010
      
1
100%
12
1.08
13,090
         
Analyst Programmer /Data Manager
NOB
2,655
      
1
25%
12
1.08
8,602
           
 Data Management Assistant
GS3
666
         
2
50%
12
1.08
8,631
           
Asst. Coordination Manager
NOA
2,224
      
1
50%
12
1.08
14,412
         
Administrative Officer
GS5
1,010
      
1
100%
12
1.08
13,090
         
Assistant Programme Manager
NOA
2,224
      
1
50%
12
1.08
14,412
         
Sr. Field Attendant
GS2
553
         
2
100%
12
1.08
14,334
         
Field Attendant /FA
GS1
499
         
3
100%
12
1.08
19,401
         
Subtotal 
       447,438 
Consultant
 $ Rate 
 No. 
CCD Consultant (for Emerging infectious diseases)
     24,506 
         1 
         24,506 
        -   
                - 
Subtotal 
         24,506 
Travel and Perdiem
 $ Rate 
 No. 
Local Transport including hiring vehicle
 bulk 
10,500
$   
1
10,500
         
ICDDR,B Transport
14,280
$   
1
14,280
         
Perdiem and Lodging
3,000
$    
1
3,000
           
International Travel with perdiem
9,600
$    
1
10,400
         
Subtotal 
         38,180 
Supplies:
 $ Rate 
 No. 
Supplies - stock/nonstock:
Office Supplies Stock/Non Stock
600
$       
12
7,200
           
Specimen collection supplies - reagents
-
              
Supplies - Lab reagents
1,200
$    
12
14,400
         
PPE and other precautionary measures for Covid
15,200
         
Liquid Nitrogen Supplies
-
              
Dry ice for specimen transport
-
              
Office maintenance, cleaning and general supplies 
-
              
Supplies capitals:
Name of the items
unit
Rate
Qty.
Total
Laptop/Docking Computer with all Accessories
2,200
      
1
2,200
   
Air Condition for office
1,250
      
1
1,250
   
Refrigerator
1,500
      
1
1,500
   
IPS
1,000
      
1
1,000
   
Subtotal Capital supplies:
5,950
           
Subtotal 
         42,750 
Others
 $ Rate 
 No. 
Staff Development Training (Int. and local)
500
$       
1
         
              500 
Workshop / Seminar (Int. and local)
500
$       
1
         
              500 
Project implementation meeting and workshop
500
$       
1
         
              500 
Other services, Stipend / Labor charge
48
$         
12
       
              576 
Communication( Fax, Phone bill, Courier, postage etc.)
209
$       
12
       
           2,502 
Shipment and Int. Courier
8,000
$    
2
         
         16,000 
Office set up, Rent, Utilities, Repairs & Maintenance
1,394
$    
1
         
           1,394 
Lab Test Cost
6,500
$    
1
         
           6,500 
Printing and photocopy
153
$       
12
       
           1,836 
Subtotal
         30,308 
Contractual
 $ Rate 
 No. 
To be named
3,714
      
1
         
           3,714 
-
          
-
      
                -   
Subtotal
           3,714 
Equipment (Capital Items valued >/=US$5000)
 $ Rate 
 No. 
Photocopy Machine
5,000
      
-
      
                -   
Other Capital items
-
          
-
      
                -   
Subtotal
                - 
Total Project cost
       586,896 
Total Project Admin and support cost (PAS):
       174,938 
PEI Admin Cost (11%)
         64,559 
icddr,b support cost: (19%)
       110,380 
Total Budget:
761,834
       

 
47 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
 
Budget Justifications 
Please provide one page statement justifying the budgeted amount for each major item, including the use of human resources, major equipment, and 
laboratory services. 
 
The study involves recruitment and training of staff, identification of all eligible vaccine recipients from 
the selected areas, vaccination, collection of blood and stool samples, and follow up of the vaccine 
recipients. 
 
INVESTIGATORS AND STAFF  
The amount budgeted for the investigators reflect a reasonable estimate of the minimum time required to 
implement the study. Dr. K. Zaman will be responsible for overall implementation of the study. Dr. Md 
Yunus will provide support for overall implementation as well as advice on epidemiological aspects. 
• Research Investigator: Research investigator will coordinate all field research activities and will 
be responsible overall field management. 
• Senior Scientific Officer: Will be responsible for overall lab activities of the study. 
• Senior Field Research Officer: For study support and maintaining all documents.  
• Field Research Officer:  For supervising all field activities. 
• Field Research supervisor: For supervising all field activities. 
• Field Research Assistant: Informs families of the purpose and activities of the study, offer 
participation, complete different documents, etc. 
• Female Field Worker/Surveillance Worker: Informs families of the purpose and activities of the 
study, offer participation, bring vaccine recipients to study clinic, complete different documents, 
etc. 
• Medical officer: Will be responsible for examination of the vaccine recipients, take written 
consent, collect blood and other samples, provide necessary treatment, record and fill up of serious 
adverse events form and preparation of reports.  
• Nurse: Assist medical officer in examining the patients, responsible for vaccination, dispensing 
medicines, collection of samples and filling of forms.  
• Senior Research officer (Lab): For processing of samples, maintaining cold chain, and 
accountability of vaccines. 
• Research Officer (Lab): For processing and shipping samples. 
• Analyst Programmer / Senior Programmer: Overall data management and designing systems for 
entry and analysis of data. 
• Senior Data Management Officer: Data entry and assist Senior Programmer. 
• Senior Data Management assistant: Data entry and assist Senior Programmer. 
• Assistant Coordination Manager: For managing all project related financial and administrative 
activities. 
• Assistant Programme Manager: Responsible for project management and administrative activities 
• Sr. Field Attendant & Field Attendant: To support study related activities 
 
TRAVEL COSTS  
• International travel - Travel costs related to attending meetings and presenting findings of the 
study at conferences. 

 
48 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
• icddr,b transport - Transport costs for regular field work, monitoring data collection 
methods/techniques, schedule family visits, vaccine recipients transportation cost to/from the 
study clinic, office. 
• Local transportation including hiring vehicle charges and guest house costs - Costs for land 
transport, vehicle hiring charges, guest house costs. 
 
SUPPLIES AND OTHER COSTS 
• Office Supplies Stock/Non Stock - This includes office and field related stock/non-stock supplies, 
housekeeping, janitorial, stationary, book & periodicals, food & beverage, cabinets, furniture, 
electronics and electric goods, mobile phone set, gasoline, petrol, diesel, fuel, CNG, tools, spares 
and servicing and other procurement and material item cost etc. for field activities. 
 
Other Support 
Describe sources, amount, duration, and grant number of all other research funding currently granted to PI or under consideration.  
 
No other support will be provided for this study. 
 
 
 
 

 
49 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Biography of the Investigators 
 
1.  Name: K. Zaman 
 
   Present Position: Senior Scientist and Epidemiologist, icddr,b 
 
   Educational background: (last degree and diploma & training relevant to the present research  
    proposal) 
 
 
Institution 
Year 
PhD 
Johns Hopkins Bloomberg School of Public Health, USA 
1999 
MPH 
Johns Hopkins Bloomberg School of Public Health, USA 
1992 
MBBS 
Rajshahi Medical College, Bangladesh 
1978 
FRCP 
Royal College of Physicians of Edinburgh 
2020 
 
 
    Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
PHRP 
2854414 
 Taken on       
07/13/2020 
 
 
Note: If the response is “no”, please get certification from CITI or NIH before study 
initiation and submit a copy to the Committee Coordination Secretariat 
 
   List of ongoing research protocols/ activities   
        
Protocol/ 
Activity  
Number 
Role in the protocol/ 
activity (PI, Co-PI, Co-I)  
Starting date 
 
End date 
Percentage of 
time 
PR 15050 
PI 
July 2015 
Sep 2021 
5 
PR 16014 
PI 
March 2016 
Dec 2019 
5 
PR 14059 
PI 
Jan 2018 
Dec 2020 
5 
PR 18045 
PI 
Sep 2018 
Sep 2020 
20 
PR 18050 
PI 
Sep 2018 
Sep 2020 
10 
PR 18062 
PI 
Sep 2018 
April 2020 
20 
 
   
Publications  
 
Types of publications 
Numbers 
a. Original scientific papers in peer-review journals                                
201 
b.   Peer reviewed articles and book chapters                                                    3 
c. Papers in conference proceedings 
135 
d. Letters, editorials, annotations, and abstracts in peer-reviewed journals   4 
e. Working papers 
 
f. Monographs 
 
 
 
    

 
50 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Five recent publications including publications relevant to the present research protocol 
 
1. Zaman K, Anand A. Sequential inactivated and oral polio vaccine schedules: a balancing act.  
Lancet ID 2020 (in press). 
 
2. Zaman K, Kingma R, Yunus M, van Straaten I, Mekkes D, Bouwstra X, Gunale B, 
Kulkarni PS. Safety, immunogenicity and lot-to-lot consistency of a new Bivalent Oral Polio 
Vaccine (bOPV) in healthy Infants: Results of a Phase III, observer blind, randomized, 
controlled clinical study. Vaccine 2019 Jun 22. doi:10.1016/j. vaccine.2019.06.048 [Epub 
ahead of print]. 
 
3. Zaman K, Anand A. Complex tasks to estimate immune responses to various poliovirus 
vaccines and vaccination schedules. Lancet ID 2019 Jul 23. pii: S1473-3099(19)30322-6. 
doi: 10.1016/S1473-3099(19)30322-6. [Epub ahead of print].  
 
4. Zaman K, Estívariz CF, Morales M, Yunus M, Snider CJ,  Gary HE Jr, Weldon WC, 
Oberste MS, Wassilak SG, Pallansch MA, Anand A. Immunogenicity of type 2 monovalent 
oral and inactivated Poliovirus vaccines for type 2 poliovirus outbreak response: An open -
level randomized controlled trial. Lancet ID 2018 Jun;18(6):657-665. 
 
5. Zaman K, Anh DD, Victor JC, Shin S, Yunus M, Dallas MJ, Podder G, Thiem VD, Mai LP,   Luby 
SP, Coia ML, Lewis K, Rivers SJ, Sack DA, Clemens JD, Scodel F, Steele AD, Neuzil  KM, Ciarlet 
M. Efficacy of the pentavalent rotavirus vaccine against sever rotavirus among infants in developing 
countries in Asia: A randomized, double-blind placebo-controlled trial. Lancet 2010 Aug 
21;376(9741): 615-23. Epub 2010 Aug 6. 
 
                     

 
51 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
-------------------------------------------------------------------------------------------------------------------------- 
Name: Md. Yunus 
Present Position: Emeritus Scientist 
 
Educational background: (last degree and diploma & training relevant to the present research proposal) 
 
 
Institution 
Year 
Degree 
MBBS 
Dhaka Medical College, Dhaka University 
1968 
Degree 
MSc. CHDC 
London School of Hygiene & Tropical Medicine, London 
 
1982 
Diploma 
 
 
Training 
 
 
Training 
 
 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No  
  
 
Yes  
 
NIH 
2361583 
29/03/2017 (date of 
completion) 
 
Note: If the response is “no”, please get certification from CITI or NIH before study initiation and 
submit a copy to the Committee Coordination Secretariat 
 
List of ongoing research protocols/ activities   
        
Protocol/ Activity  
Number 
Role in the protocol/ activity 
(PI, Co-PI, Co-I)  
Starting date 
 
End date 
Percentage of 
time 
PR # 17034 
Co-I 
Oct 2017 
Dec 2018 
20 
PR # 15016 
Co-I 
Jan 2017 
Jan 2019 
5 
PR # 16077 
Co-I 
Dec 2016 
Dec 2018 
10 
PR # 16014 
Co-I 
Apr 2016 
Mar 2020 
5 
PR # 15111 
PI 
Feb 2016 
Aug 2020 
20 
PR# 15050 
Co-I 
Jul 2015 
Jul 2018 
3 
PR # 15007 
Co-PI 
Feb 2015 
Jan 2018 
10 
RRC PR#16014 
 
15/03/16 
31/12/19 
5 % 
5 
 
 
 
Publications  
 
Types of publications 
Numbers 
Original scientific papers in peer-review journals                                
238 
  Peer reviewed articles and book chapters                                                                
8 
Papers in conference proceedings 
101 
Letters, editorials, annotations, and abstracts in peer-reviewed journals   
15 
Working papers 
6 
Monographs 
0 
 
Five recent publications including publications relevant to the present research protocol 

 
52 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
  
1. 
Schwartz, L. M., Zaman, K., Yunus, M., Basunia, A. H., Faruque, A. S. G., Ahmed, T., . . . Victor, 
J. C. (2019). Impact of rotavirus vaccine introduction in children less than 2 years of age 
presenting for medical care with diarrhea in rural Matlab, Bangladesh. Clin Infect Dis. doi: 
10.1093/cid/ciz133  
2. 
Rahman, M., Sohel, N., Yunus, F. M., Alam, N., Nahar, Q., Streatfield, P. K., & Yunus, M. 
(2019). Arsenic exposure and young adult's mortality risk: A 13-year follow-up study in Matlab, 
Bangladesh. Environ Int, 123, 358-367. doi: 10.1016/j.envint.2018.12.006 
3. 
Zman K, Estivariz CF.,Morales M, Yunus M, Snider CJ, Gary Jr HE, Weldon WC, Oberste MS, Wassilak 
SG, Pallansch M A & Anand A. 2018a. Immunogenicity of type 2 monovalent oral and inactivated 
poliovirus vaccines for type 2 poliovirus outbreak response: an open-label, randomised controlled trial. 
Lancet Infect Dis, 18, 657-665 
4. 
Anand A, Zaman K, Estívariz CF, Yunus M, Gary HE, Weldon WC, Bari TI, Steven Oberste M, Wassilak 
SG, Luby SP, Heffelfinger JD, Pallansch MA.Early priming with inactivated poliovirus vaccine (IPV) and 
intradermal fractional dose IPV administered by a microneedle device: A randomized controlled 
trial.Vaccine. 2015 Nov 27;33(48):6816-22. doi: 10.1016/j.vaccine.2015.09.039.PMID:26476367 
5. 
Yunus M, Sohel N, Hore SK, Rahman M.Arsenic exposure and adverse health effects: a review of recent 
findings from arsenic and health studies in Matlab, Bangladesh.Kaohsiung J Med Sci. 2011 
Sep;27(9):371-6. Epub 2011 Jul 6. 
 
 
 
------------------------------------------------------------------------------------------------------------------------------- 
 
 

 
53 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Name: Abhijeet Anand 
Present position: Epidemiologist, Polio Eradication Branch, Global Immunization Division, CDC 
Education background: 
 
Institution 
Year 
MBBS 
Byramjee Jeejeebuoy Medical College, Pune, 
India 
2001 
MPH (Epidemiology) 
Epidemiology, Johns Hopkins University 
Bloomberg School of Public Health 
2003 
Epidemic Intelligence Service 
(EIS) 
US Centers for Disease Control and 
Prevention 
2005 
 
 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
277837334 
20 Dec 
2021 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
  13 
Letters, editorials in peer-reviewed journals   
8 
 
Recent publications including publications relevant to the present research protocol: 
1. Anand A, Luman ET, O’Connor P. Building on Success – Potential to improve coverage of multiple health 
interventions in Africa through integrated delivery with routine childhood vaccination. (Accepted: The 
Journal of Infectious Diseases) 
2. Wiesen E, Wannemuehler K, Goodson JL, Anand A, Mach O, Thapa A, O'Connor P, Linayage J, Diorditsa 
S, Hasan AS, Uzzaman S, JalilMondal MD. Stability of the age distribution of measles cases over time 
during outbreaks in Bangladesh, 2004-2006. J Infect Dis. 2011 Jul;204Suppl 1:S414-20. 
3. Upreti SR, Thapa K, Pradhan YV, Shakya G, Sapkota YD, Anand A, Taylor T, Mach O, Reef S, 
Pattamadilok S, Liyanage J, O'Connor P, Sedai T, Bhandary SR, Partridge J, Schluter W. Developing rubella 
vaccination policy in Nepal--results from rubella surveillance and seroprevalence and congenital rubella 
syndrome studies. J Infect Dis. 2011 Jul;204Suppl 1:S433-8. 
4. O'Connor PM, Liyanage JB, Mach O, Anand A, Ramamurty N, Balakrishnan MR, Singh S. South-East Asia 
Regional update on measles mortality reduction and elimination, 2003-2008. J Infect Dis. 2011 Jul;204 Suppl 
1:S396-402. 
5. Anand A, Shiraishi RW, Bunnell RE, Jacobs K, Solehdin N, Abdul-Quader AS, Marum LH, Muttunga JN, 
Kamoto K, Aberle-Grasse JM, Diaz T. Knowledge of HIV status, sexual risk behaviors and contraceptive 
need among people living with HIV in Kenya and Malawi. AIDS. Jul 31 2009;23(12):1565-1573. 
 
 
------------------------------------------------------------------------------------------------------------------------------- 
 
 

 
54 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Name: Cindi Snider 
Present position: Epidemiologist, Polio Eradication Branch, Global Immunization Division, CDC 
Education background: 
 
Institution 
Year 
BSPH 
University of North Carolina at Chapel Hill 
1997 
MHS (Epidemiology) 
Johns Hopkins University Bloomberg School 
of Public Health 
2001 
PhD (Epidemiology) 
University of North Carolina at Chapel Hill 
2011 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
227211624 
25- Nov-2021 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
 11 
Book chapters and review articles                                                           
2 
 
Recent publications including publications relevant to the present research protocol: 
1. Snider CJ, Zaman K, Estivariz CF, Yunus M, Weldon WC, Wannemuehler KA, Oberste MS, Pallansch MA, 
Wassilak SG, Bari TIA, Anand A. Immunogenicity of full and fractional dose of inactivated poliovirus 
vaccine for use in routine immunisation and outbreak response: an open-label, randomised controlled trial. 
Lancet. 2019 Jun 29;393(10191):2624-2634. doi: 10.1016/S0140-6736(19)30503-3. Epub 2019 May 16. 
2. Farag NH, Mansour Z, Torossian L, Said R, Snider CJ, Ehrhardt D. Feasibility of jet injector use during 
inactivated poliovirus vaccine house-to-house vaccination campaigns. Vaccine. 2018 Aug 6;36(32 Pt 
B):4935-4938. doi: 10.1016/j.vaccine.2018.06.011. 
3. Zaman K, Estívariz CF, Morales M, Yunus M, Snider CJ, Gary HE Jr, Weldon WC, Oberste MS, Wassilak 
SG, Pallansch MA, Anand A. Immunogenicity of type 2 monovalent oral and inactivated poliovirus vaccines 
for type 2 poliovirus outbreak response: an open-label, randomised controlled trial. Lancet Infect Dis. 2018 
Jun;18(6):657-665. doi: 10.1016/S1473-3099(18)30113-0. Epub 2018 Mar 20.  
4. Estivariz CF, Snider CJ, Anand A, Hampton LM, Bari TI, Billah MM, Chai SJ, Wassilak SG, Heffelfinger 
JD, Zaman K. Lessons Learned From the Introduction of Inactivated Poliovirus Vaccine in Bangladesh. J 
Infect Dis. 2017 Jul 1;216(suppl_1):S122-S129 
5. Billah MM, Zaman K, Estivariz CF, Snider CJ, Anand A, Hampton LM, Bari TIA, Russell KL, Chai SJ. 
Cold-Chain Adaptability During Introduction of Inactivated Polio Vaccine in Bangladesh, 2015. J Infect Dis. 
2017 Jul 1;216(suppl_1):S114-S121 
 
------------------------------------------------------------------------------------------------------------------------------- 
 
 

 
55 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Name: Concepcion F. Estivariz 
Present position: Epidemiologist, Polio Eradication Branch, Global Immunization Division, CDC 
Education background: 
 
Institution 
Year 
MD 
Universidad Autonoma, Faculty of Medicine, 
Madrid, Spain   
1990 
Physician Specialist 
Physician Specialist in Endocrinology and 
Nutrition, Ministry of Health, Spain 
1995 
Epidemic Intelligence 
Service (EIS) 
US Centers for Disease Control and Prevention 
2003 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
29524155 
26- Nov-2021 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
  38 
Book chapters and review articles                                                           
7 
Abstracts in scientific meetings    
32 
 
Recent publications including publications relevant to the present research protocol: 
1. Estivariz CF, Snider CJ, Anand A, et al. Lessons Learned From the Introduction of Inactivated Poliovirus 
Vaccine in Bangladesh. J Infect Dis 2017;216:S122-s129 
2. Anand, A., K. Zaman, Estivariz CF, Yunus M, Gary HG, Weldon WC, Oberste MS, Wassilak SG, Luby SP, 
Heffelfinger JD, Pallansch MA. "Early priming with inactivated poliovirus vaccine (IPV) and intradermal 
fractional dose IPV administered by a microneedle device: A randomized controlled trial." Vaccine 2016, 33: 
6816-6822. 
3. Cardemil CV, Estivariz C, Shrestha C, Sherchand JB, Sharma A, Gary HE,  Oberste MSJr, Weldon WC 3rd,  
Bowen MD,  Vinje J, Schluter WW, Anand A, Mach O,Chu SY. "The effect of diarrheal disease on bivalent 
oral polio vaccine (bOPV) immune response in infants in Nepal." Vaccine 2016, 34(22): 2519-2526. 
4. Emperador, D. M., Velasquez DE, Estivariz CF, Lopman B, Jiang B, Parashar U, Anand A., Zaman K. 
"Interference of Monovalent, Bivalent, and Trivalent Oral Poliovirus Vaccines on Monovalent Rotavirus 
Vaccine Immunogenicity in Rural Bangladesh." Clinical Infectious Diseases 2016, 62(2): 150-156. 
5. Estívariz CF, Anand A, Gary HE Jr, Rahman M, Islam J, Bari TI, Wassilak SGF, Chu SY, Weldon WC, 
Pallansch MA, Heffelfinger JD, Luby SP, Zaman K.  Immunogenicity of three doses of bivalent, trivalent, or 
type-1 monovalent oral poliovirus vaccines with a 2 week interval between doses in Bangladesh: an open-
label, noninferiority, randomised, controlled trial.  Lancet ID, 2015, 5(8): 898-904.   
 
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56 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Name: Amanda L. Wilkinson  
Present position: Epidemiologist, Polio Eradication Branch, Global Immunization Division, CDC 
Education background:  
 
 
Institution 
Year 
BS 
Pennsylvania State University  
2010 
PhD (Nutritional Sciences, 
Epidemiology concentration) 
Cornell University   
2015 
Epidemic Intelligence Service 
(EIS) 
US Centers for Disease Control and 
Prevention 
2018 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
20370998 
26-Jun-2021 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
7 
Book chapters and review articles                                                           
2 
Abstracts in scientific meetings    
12 
 
Recent publications including publications relevant to the present research protocol: 
1. Kalkowska DA, Pallansch MA, Wilkinson AL,  Bandyopadhyay AS, Konopka‐Anstadt JL, Burns CC, 
Oberste MS, Wassilak SGF, Badizadegan K, Thompson KM. Updated Characterization of Outbreak 
Response Strategies for 2019–2029: Impacts of Using a Novel Type 2 Oral Poliovirus Vaccine Strain. Risk 
Analysis. 2020. https://doi.org/10.1111/risa.13622.  
2. Cunningham SA, Shaikh NI, Nhacolo A, Raghunathan PL, Kotloff K, Naser AM, Mengesha MM, Adedini 
SA, Misore T, Onuwchekwa UU, Worrell MC, El Arifeen S, Assefa N, Chowdhury A, Kaiser R, Madhi S, 
Mehta A, Obor D, Sacoor C, Sow A, Tapia M, Wilkinson A, Breiman R, Child Health and Mortality 
Prevention Surveillance (CHAMPS) Methods Consortium. Health and Demographic Surveillance Systems 
Within the Child Health and Mortality Prevention Surveillance Network. Clinical Infectious Diseases. 2019 
Oct 9;69(Supplement_4):S274-9. Cited by 6.  
3. Wilkinson AL, Pedersen SH, Urassa M, Michael D, Andreasen A, Todd J, Kinung'hi SM, Changalucha J, 
McDermid JM. Maternal systemic or cord blood inflammation is associated with birth anthropometry in a 
Tanzanian prospective cohort. Tropical Medicine & International Health, 2017; 22(1):52-62. Cited by 13.    
4. Pedersen SH, Wilkinson AL, Andreasen A, Warhurst DC, Kinunghi S, Urassa M, Mkwashapi D, Todd J, 
Changalucha J, McDermid JM.  Longitudinal analysis of mature breast milk and serum immune composition 
among mixed HIV-status mothers and their infants. Clinical Nutrition, 2016; 35(4):871-9. Cited by 18.   
5. Wilkinson AL, Pedersen SH, Urassa M, Mkwashapi D, Kinunghi S, Todd J, Changalucha J, McDermid JM. 
Associations between gestational anthropometry, maternal HIV, and fetal and early infancy growth in a 
prospective rural/semi-rural Tanzanian cohort, 2012–13. BMC Pregnancy and Childbirth, 2015;15(1):277. 
Cited by 15.   
 
 
 
 

 
57 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
------------------------------------------------------------------------------------------------------------------------------- 
Name: Qian An 
Present position: Mathematical Statistician, Strategic Information and Workforce Development Branch, Global 
Immunization Division, CDC 
Education background: 
 
Institution 
Year 
BE  
Beijing Jiaotong University, Beijing, China 
1999 
MS (Biostatistics) 
University of Minnesota, Twin Cities 
2005 
PhD (Biostatistics) 
Emory University 
2014 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
29726253 
18- Dec-2021 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
37 
Abstracts in scientific meetings                                                           
14 
 
Recent publications including publications relevant to the present research protocol: 
 
1. An Q, Song R, Sionean C, Finlayson T, Wejnert C. An Innovative Approach to Assess Similarity Between 
Sex Partners. AIDS and Behavior (2018). https://doi.org/10.1007/s10461-018-2285-0.  
2. An Q, Wejnert C, Bernstein K, Paz-Bailey G. Syphilis Screening and Diagnosis Among Men Who Have 
Sex With Men, 2008-2014, 20 U.S. Cities. JAIDS Journal of Acquired Immune Deficiency Syndromes 
2017. 75(S3):363–369. DOI: 10.1097/QAI.0000000000001412. 
3. An Q, Song R, Finlayson TJ, Wejnert C, Paz-Bailey G. Estimated HIV Inter-test Interval Among Persons 
at High Risk for HIV Infection In the U.S. Am J Prev Med. 2017; 53(3):355–362.  
4. An Q, Kang J, Song R, Hall HI. A Bayesian Hierarchical model with novel prior specifications for 
estimating HIV testing rates. Statistics in Medicine. 2016; 35(9): 1471-1487. DOI: 10.1002/sim.6795. 
5. An Q, Chronister K, Song R, Yang B, Pearson M, Chan S and etc. Agreement of self-report and medical 
records review data on HIV testing behavior. Annals of Epidemiology. 2016; 26(4): 255-260.   
 
 
 

 
58 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
------------------------------------------------------------------------------------------------------------------------------- 
Name: Steven Wassilak 
Present position: Senior Epidemiologist, Polio Eradication Branch, Global Immunization Division, CDC 
Education background: 
 
Institution 
Year 
BA (Biology) 
University of Missouri-St. Louis 
1973 
MD 
St Louis University School of Medicine 
1977 
Epidemic Intelligence Service 
(EIS) 
US Centers for Disease Control and Prevention 
1980 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
19789 
 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
  45 
Book chapters and review articles                                                           
14 
Letters, editorials in peer-reviewed journals   
18 
 
Recent publications including publications relevant to the present research protocol: 
1. Wassilak S, Pate MA, Wannemuehler K, et al. Outbreak of type 2 vaccine-derived poliovirus in Nigeria: 
emergence and widespread circulation in an underimmunized population. J Infect Dis 2011;203:898-909.  
2. Kidd S.  Goodson JL.  Aramburu J.  Morais A.  Gaye A.  Wannemuehler K.  Buffington J.  Gerber S.  
Wassilak S.  Uzicanin A. Poliomyelitis outbreaks in Angola genetically linked to India: risk factors and 
implications for prevention of outbreaks due to wild poliovirus importations.  Vaccine.  29(21):3760-6, 2011 
May 12.  
3. Economic analysis of the global polio eradication initiative. DuintjerTebbens RJ.  Pallansch MA.  Cochi SL.  
Wassilak SG.  Linkins J.  Sutter RW.  Aylward RB.  Thompson KM. Vaccine.  29(2):334-43, 2010 Dec 16.  
4. Challenges faced by the global polio eradication initiative. Wassilak S.  Orenstein W. Expert Review of 
Vaccines.  9(5):447-9, 2010 May.  
5. Kojouharova M, Zuber PL, Gyurova S, Fiore L, Buttinelli G, Kunchev A, Vladimirova N, Korsun N, 
Filipova R, Boneva R, Gavrilin E, Deshpande JM, Oblapenko G, Wassilak SG: Importation and circulation 
of poliovirus in Bulgaria in 2001. Bull World Health Organ 2003;81(7):476-81. 
 
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59 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Name: Mark A. Pallansch 
Present position: Director, Division of Viral Diseases, National Center for Immunization and Respiratory 
Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 
 
 
Education background: 
 
Institution 
Year 
BS (Biochemistry) 
Virginia Tech   Blacksburg, VA 
1976 
PhD (Biochemistry) 
University of Wisconsin – Madison  , WI 
1982 
Virology 
Rockefeller University New York, NY 
1984 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
7660 
 
 
Publications: 
Type of publication 
       Number 
Original scientific papers in peer-review journals                                
 264 
Book chapters and review articles                                                           
36 
Abstracts in scientific meetings    
 133 
 
Recent publications including publications relevant to the present research protocol: 
1. Snider CJ, Zaman K, Estivariz CF, Yunus M, Weldon WC, Wannemuehler KA, Oberste MS, Pallansch MA, 
Wassilak SGF, Bari TIA, Anand A.  Immunogenicity of full and fractional dose of inactivated poliovirus 
vaccine for use in routine immunisation and outbreak response: an open-label, randomised controlled trial.  
Lancet. 2019 Jun 29;393(10191):2624-2634. 
2. Zaman K, Estívariz CF, Morales M, Yunus M, Snider CJ, Gary HE Jr, Weldon WC, Oberste MS, 
Wassilak SG, Pallansch MA, Anand A.  Immunogenicity of type 2 monovalent oral and inactivated 
poliovirus vaccines for type 2 poliovirus outbreak response: an open-label, randomised controlled 
trial.  Lancet Infect Dis, 2018;18:657-665. 
3. Anand A, Molodecky NA, Pallansch MA, Sutter RW.  Immunogenicity of two doses of fractional 
intradermal inactivated poliovirus vaccine: a novel dose sparing immunization schedule.  Vaccine 
2017;35:2993-2998 
4. Anand A, Zaman K, Estivariz CF, Yunos M, Gary HE Jr, Weldon WC, Bari TI, Oberste MS, 
Wassilak SG, Luby SP, Heffelfinger JD, Pallansch MA. Early priming with inactivated poliovirus 
vaccine (IPV) and intradermal fractional dose IPV administered by a microneedle device: a 
randomized controlled trial.  Vaccine 2015;33:6816-22 
5. Estívariz CF, Jafari H, Sutter RW, John TJ, Jain V, Agarwal A, Verma H, Pallansch MA, Singh AP, 
Guirguis S, Awale J, Burton A, Bahl S, Chatterjee A, Aylward RB.  Immunogenicity of supplemental doses 
of poliovirus vaccine for children age 6–9 months in Moradabad, India: a community-based, randomised 
controlled trial.  Lancet Infectious Diseases 2015;15:898-904.  
 
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60 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Name: M. Steve Oberste 
Present position: Chief, Polio and Picornavirus Laboratory Branch, Division of Viral Diseases, CDC 
Education background: 
 
Institution 
Year 
BS (Genetics) 
College of Letters and Sciences, University of 
California at Davis; Davis, California 
1982 
PhD (Immunology 
and Medical 
Microbiology) 
College of Medicine, University of Florida; 
Gainesville, Florida 
1988 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
8333 
 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
  87 
Letters, editorials in peer-reviewed journals   
7 
 
Recent publications including publications relevant to the present research protocol: 
1. Anand A, Zaman K, Estivariz C, Yunus M, Gary H, Weldon WC, Bari TI, Oberste MS, Wassilak S, 
Luby S, Heffelfinger JD, Pallansch MA. 2015. Early priming with inactivated poliovirus vaccine (IPV) 
and intradermal fractional dose IPV administered by a microneedle device: A randomized controlled 
trial. Vaccine 33:6816-6822. (PMID: 26476367) 
2. Sáez-Llorens X, Clemens R, LeRoux Roels G, Jimeno, J, Costa Clemens SA, Weldon WC, Oberste MS, 
Molina N, Bandyopadhyay AS. 2016. Immunogenicity and safety of a novel monovalent high-dose 
inactivated poliovirus type 2 vaccine in infants: a comparative, observer-blind, randomised controlled 
study. Lancet Infect Dis 16:321-330. (PMID: 26719058) 
3. Asturias EJ, Bandyopadhyay AS, Self S, Rivera L, Saez-Llorens X, Lopez E, Melgar M, Gaensbauer JT, 
Weldon WC, Oberste MS, Borate BR, Gast C, Clemens R, Orenstein W, O’Ryan G M, Jimeno J,  Costa 
Clemens SA, Ward J, Rüttimann R. 2016. Humoral and intestinal immunity induced by new schedules of 
bivalent OPV and one or two doses of IPV in an open, randomised, controlled trial in Latin American 
infants. Lancet 388:158-169. (PMID: 27212429) 
4. Habib MA, Soofi S, Mach O, Samejo T, Alam D, Bhatti Z, Weldon WC, Oberste MS, Sutter R, Bhutta 
ZA. Effect of booster doses of poliovirus vaccine in previously vaccinated children, clinical trial results 
2013. Vaccine 34:3803-3809. (PMID: 27269054) 
5. Brickley EB, Strauch C, Wieland-Alter W, Connor RI, Lin S, Weiner JA, Ackerman ME, Arita M, 
Weldon WC, Oberste MS, Sáez-Llorens X, Bandyopadhyay AS, Wright PF. 2018. Intestinal immune 
responses to type 2 oral polio vaccine challenge in infants previously immunized with bOPV and either 
high-dose or standard IPV. J Infect Dis In Press. (PMID: 29304199) 
 
 
 
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61 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Name: Jacquelyn Lickness 
Present position: Public Health Analyst, Polio Eradication Branch, Global Immunization Division, U.S. CDC 
Education background: 
Degree 
Institution 
Year 
MPH 
Emory University 
2014 
 
 
 
 
 
 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
18892 
 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
 2 
Letters, editorials in peer-reviewed journals   
 
 
Recent publications including publications relevant to the present research protocol: 
 
1. Lickness JS, Gardner T, Diop OM, et al. Surveillance to Track Progress Toward Polio Eradication – 
Worldwide, 2018-2019. MMWR Morb Martal Wkly Rep 2020;69:623-629.  
2. Bartz FE, Lickness JS, Heredia N, Fabiszewski de Aceituno A, Newman KL, Hodge DW, Jaykus L-A, 
García S, Leon JS. 2017. Contamination of fresh produce by microbial indicators on farms and in 
packing facilities: elucidation of environmental routes. Appl Environ Microbiol 83:e02984-16. 
https://doi.org/10.1128/AEM.02984-16.  
 
 
 

 
62 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
------------------------------------------------------------------------------------------------------------------------------- 
Name: Jaymin Patel 
Present position: Global Immunization Division, CDC, USA 
Education background: 
Degree 
Institution 
Year 
PhD – Epidemiology 
University of North Carolina at Chapel Hill 
2016 
MPH – Global Epidemiology 
Emory University 
2011 
BS–Biology; BA–Env. Studies 
University of North Carolina at Chapel Hill 
2008 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
9300 
 
 
Publications: 
Types of publications 
Numbers 
Original scientific papers in peer-review journals                                
20 
Papers in conference proceedings 
13 
 
Five recent publications including publications relevant to the present research protocol: 
1. Patel JC, Soeters HM, Diallo AO, et al. MenAfriNet: A Network Supporting Case-Based Meningitis 
Surveillance and Vaccine Evaluation in the Meningitis Belt of Africa. The Journal of infectious diseases 2019; 
220(Supplement_4): S148-s54. 
2. Patel JC, Diop OM, Gardner T, et al. Surveillance to Track Progress Toward Polio Eradication - Worldwide, 
2017-2018. MMWR Morb Mortal Wkly Rep 2019; 68(13): 312-8. 
3. Patel JC, George J, Vuong J, Potts CC, Bozio C, Clark TA, et al. Rapid Laboratory Identification of Neisseria 
meningitidis Serogroup C as the Cause of an Outbreak - Liberia, 2017. MMWR Morb Mortal Wkly Rep. 2017 
Oct 27;66(42):1144-7. 
4. Patel JC, Hathaway NJ, Parobek CM, Thwai KL, Madanitsa M, Khairallah C, et al. Increased risk of low 
birth weight in women with placental malaria associated with P. falciparum VAR2CSA clade. Sci Rep. 2017 
Aug 11;7(1):7768. 
5. Patel JC, Mwapasa V, Kalilani L, et al. Absence of Association Between Sickle Trait Hemoglobin and 
Placental Malaria Outcomes. The American journal of tropical medicine and hygiene 2016; 94(5): 1002-7. 
 
 

 
63 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
------------------------------------------------------------------------------------------------------------------------------- 
Name: Jennifer Anstadt 
Present position: Lead Microbiologist, Vaccine Development Laboratory, Polio and Picornavirus Laboratory 
Branch, Division of Viral Diseases, Centers for Disease Control and Prevention 
Education background: 
 
Institution 
Year 
BS (Biology) 
Fairfield University  
2000 
PhD (Microbiology & Immunology) 
University of North Carolina, Chapel Hill 
2007 
Postdoctoral Fellow (Pediatric 
Infectious Diseases) 
Vanderbilt University School of Medicine 
2012 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
29741394 
12 Dec 2021 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
  16 
Letters, editorials in peer-reviewed journals   
2 
 
Recent publications including publications relevant to the present research protocol: 
1. Sáez-Llorens X, Bandyopadhyay AS, Gast C, De Leon T, DeAntonio R, Jimeno J, Caballero MI, Aguirre 
G, Oberste MS, Weldon WC, Konopka-Anstadt JL, Modlin J,  Bachtiar NS, Fix A, Konz J, Clemens R, 
Costa-Clemens SA, Rüttimann R. Safety and immunogenicity of two novel live attenuated oral poliovirus 
type 2 vaccines candidates in healthy children and infants: two randomised trials. (Under review, Lancet) 
2. De Coster I, Leroux-Roels I, Bandyopadhyay AS, Gast C, Withanage K, Steenackers K, De Smedt P, 
Aerssens A, Leroux-Roels G, Oberste MS, Konopka-Anstadt JL, Weldon WC, Fix A, Konz J, Wahid R, 
Modlin J, Clemens R, Costa-Clemens SA, Bachtiar NS, Van Damme P. An evaluation of the safety and 
immunogenicity of two novel live attenuated serotype 2 oral poliovirus vaccine candidates in healthy 
adults; a phase 2, partial blind, randomised, placebo-controlled study. (Under review, Lancet) 
3. Konopka-Anstadt JL, Campagnoli R, Vincent A, Shaw J, Wei L, Wynn NT, Smithee SE, Bujaki E, Te 
Yeh M, Laassri M, Zagorodnyaya T, Weiner AJ, Chumakov K, Andino R, Macadam A, Kew O, Burns CC. 
Development of a new oral poliovirus vaccine for the eradication end game using codon deoptimization. 
NPJ Vaccines. 2020 Mar 20;5:26. eCollection 2020. 
4. Schubert RD, Hawes IA, Ramachandran PS, Ramesh A, Crawford ED, Pak JE, Wu W, Cheung CK, 
O'Donovan BD, Tato CM, Lyden A, Tan M, Sit R, Sowa GA, Sample HA, Zorn KC, Banerji D, Khan LM, 
Bove R, Hauser SL, Gelfand AA, Johnson-Kerner BL, Nash K, Krishnamoorthy KS, Chitnis T, Ding JZ, 
McMillan HJ, Chiu CY, Briggs B, Glaser CA, Yen C, Chu V, Wadford DA, Dominguez SR, Ng TFF, 
Marine RL, Lopez AS, Nix WA, Soldatos A, Gorman MP, Benson L, Messacar K, Konopka-Anstadt JL, 
Oberste MS, DeRisi JL, Wilson MR. Pan-viral serology implicates enteroviruses in acute flaccid myelitis. 
Nat Med. 2019 Nov;25(11):1748-1752.  
5. Lopez A, Lee A, Guo A, Konopka-Anstadt JL, Nisler A, Rogers SL, Emery B, Nix WA, Oberste S, Routh 
J, Patel M. Vital Signs: Surveillance for Acute Flaccid Myelitis - United States, 2018. MMWR Morb 
Mortal Wkly Rep. 2019 Jul 12;68(27):608-614. 
6. Van Damme P, De Coster I, Bandyopadhyay AS, Revets H, Withanage K, De Smedt P, Suykens L, Oberste 
MS, Weldon WC, Costa-Clemens SA, Clemens R, Modlin J, Weiner AJ, Macadam AJ, Andino R, Kew 
OM, Konopka-Anstadt JL, Burns CC, Konz J, Wahid R, Gast C. The safety and immunogenicity of two 
novel live attenuated monovalent (serotype 2) oral poliovirus vaccines in healthy adults: a double-blind, 
single-centre phase 1 study. Lancet. 2019 Jul 13;394(10193):148-158. 
 

 
64 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
------------------------------------------------------------------------------------------------------------------------------- 
Name: Cara C. Burns 
Present position: Acting Branch Chief, Polio and Picornavirus Laboratory Branch, Division of Viral Diseases, 
CDC 
Education background: 
 
Institution 
Year 
B. S. Biochemistry 
Texas A& M University, College Station, Texas 
1985 
PhD Cellular, Viral and 
Molecular Biology 
University of Utah, Salt Lake City, Utah 
1991 
Postdoctoral training 
University of Washington Microbiology 
Department, Seattle, Washington 
1991
-
1998 
 
Ethics Certification: 
 
 
 
If Yes 
 
 
Issuing Authority 
Registration No 
Valid Until 
No 
   
Yes  
 
CDC 
30057846 
14Jan 
2022 
 
Publications: 
Type of publication 
  Number 
Original scientific papers in peer-review journals                                
 >50 
Letters, editorials in peer-reviewed journals   
0 
 
Recent publications including publications relevant to the present research protocol: 
 
1. Fagnant-Sperati CS, Ren Y, Zhou NA, Komen E, Mwangi B, Hassan J, Chepkurui A, Nzunza R, 
Nyangao J, van Zyl WB, Wolfaardt M, Matsapola PN, Ngwana FB, Jeffries-Miles S, Coulliette-Salmond 
A, Peñaranda S, Vega E, Shirai JH, Kossik AL, Beck NK, Boyle DS, Burns CC, Taylor MB, Borus P, 
Meschke JS.Fagnant-Sperati CS, et al. Validation of the bag-mediated filtration system for environmental 
surveillance of poliovirus in Nairobi, Kenya. J Appl Microbiol. 2020 Aug 2. doi: 10.1111/jam.14807. 
Online ahead of print. J Appl Microbiol. 2020. PMID: 32743931 
2. Mary M Alleman, Jaume Jorba, Sharon A Greene, Ousmane M Diop, Jane Iber, Graham Tallis, Ajay 
Goel, Eric Wiesen, Steven G F Wassilak, Cara C Burns. MMWR Morb Mortal Wkly Rep. 2020 Apr 
24;69(16):489-495. doi:10.15585/mmwr.mm6916a1. Update on Vaccine-Derived Poliovirus Outbreaks - 
Worldwide, July 2019-February 2020 PMID: 32324719 PMCID: PMC7188410 DOI: 
10.15585/mmwr.mm6916a1  
3. Zhou NA, Fagnant-Sperati CS, Komen E, Mwangi B, Mukubi J, Nyangao J, Hassan J, Chepkurui A, 
Maina C, van Zyl WB, Matsapola PN, Wolfaardt M, Ngwana FB, Jeffries-Miles S, Coulliette-Salmond 
A, Peñaranda S, Shirai JH, Kossik AL, Beck NK, Wilmouth R, Boyle DS, Burns CC, Taylor MB, Borus 
P, Meschke JS.Zhou NA, et al. Feasibility of the Bag-Mediated Filtration System for Environmental 
Surveillance of Poliovirus in Kenya. Food Environ Virol. 2020 Mar;12(1):35-47. doi: 10.1007/s12560-
019-09412-1. Epub 2019 Nov 2. Food Environ Virol. 2020. PMID: 31679104 Free PMC article.  
4. Konopka-Anstadt JL, Campagnoli R, Vincent A, Shaw J, Wei L, Wynn NT, Smithee SE, Bujaki E, Te 
Yeh M, Laassri M, Zagorodnyaya T, Weiner AJ, Chumakov K, Andino R, Macadam A, Kew O, Burns 
CC. Development of a new oral poliovirus vaccine for the eradication end game using codon 

 
65 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
deoptimization. NPJ Vaccines. 2020 Mar 20;5:26. doi: 10.1038/s41541-020-0176-7. eCollection 
2020.NPJ Vaccines. 2020. PMID: 32218998  
5. Estívariz CF, Pérez-Sánchez EE, Bahena A, Burns CC, Gary HE Jr, García-Lozano H, Rey-Benito G, 
Peñaranda S, Castillo-Montufar KV, Nava-Acosta RS, Meschke JS, Oberste MS, Lopez-Martínez I, 
Díaz-Quiñonez JA.Estívariz CF, et al. Field Performance of Two Methods for Detection of Poliovirus in 
Wastewater Samples, Mexico 2016-2017. Food Environ Virol. 2019 Dec;11(4):364-373. doi: 
10.1007/s12560-019-09399-9. Epub 2019 Sep 30. Food  Environ Virol. 2019. PMID: 31571037  
6. Van Damme P,  De Coster I,  Bandyopadhyay A, Prevets H, Withanage K, De Smedt P,  Suykens L, 
Oberste M,  Weldon W, Costa-Clemens S, Clemens R,  Modlin J, Weiner A, Macadam A, Andino R,  
Kew O, Konopka-Anstadt J, Burns CC,  Konz J, Wahid R, and Gast C. A blinded phase 1 study of the 
safety and immunogenicity of two novel live attenuated monovalent (serotype 2) oral poliovirus vaccines 
in healthy adults. Lancet, Published online June 4, 2019  http://dx.doi.org/10.1016/S0140-
6736(19)31279-6 
7. Shaw J, Jorba J, Zhao K, Iber J, Chen Q, Adu F, Adeniji A, Bukbuk D, Baba M, Henderson E, Dybdahl-
Sissoko N, Macdonald S, Weldon WC, Gumede N, Oberste MS, Kew OM, Burns CC. Dynamics of 
Evolution of Poliovirus Neutralizing Antigenic Sites and Other Capsid Functional Domains during a 
Large and Prolonged Outbreak. J Virol. 2018 Feb 14. pii: JVI.01949-17. doi: 10.1128/JVI.01949-17. 
[Epub ahead of print] PubMed PMID: 29444940; PubMed Central PMCID: PMC5899205. 
8. Montmayeur AM, Ng TF, Schmidt A, Zhao K, Magaña L, Iber J, Castro CJ, Chen Q, Henderson E, 
Ramos E, Shaw J, Tatusov RL, Dybdahl-Sissoko N, Endegue-Zanga MC, Adeniji JA, Oberste MS, 
Burns CC. High-Throughput Next-Generation Sequencing of Polioviruses. J Clin Microbiol. 2017 
Feb;55(2):606-615. doi: 10.1128/JCM.02121-16. Epub 2016 Dec 7. PMID:27927929 
9. Burns CC, Kilpatrick DR, Iber JC, Chen Q, Kew OM. Molecular Properties of Poliovirus Isolates: 
Nucleotide Sequence Analysis, Typing by PCR and Real-Time RT-PCR. Methods Mol Biol. 
2016;1387:177-212. doi: 10.1007/978-1-4939-3292-4_9.Methods Mol Biol. 2016. PMID: 26983735  
 
 
 

 
66 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Informed Consent (English and Bangla) 
  
      Information Sheet for Vaccine recipient and Sibling 
 
 
Protocol No. PR-20060 
Version No. 1.06 
Date: 03-12-2020 
                                 
 
 
 
 
 
Protocol Title: Immunogenicity of novel monovalent oral poliovirus vaccine type 2 (nOPV2) with and 
without bivalent OPV 
Investigator’s name: Dr. K. Zaman  
Organization: International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b) 
 
We would like to invite you to participate in a research project. Please read this information sheet 
carefully, and if you have any questions about the study do not hesitate to ask from the research staff. 
Feel free to discuss the project with your family or friends before you make a decision on whether to 
participate 
 
Background  
Polio disease is caused by infection with the poliovirus. Polio can cause paralysis of the legs, arms or the 
chest muscles and can result in death. There are three types of polioviruses: type 1, type 2, and type 3. To 
protect children from polio, oral poliovirus vaccines (OPV) and inactivated poliovirus vaccines (IPV) are 
used. In the EPI vaccination schedule of Bangladesh, children are given fractional IPV (fIPV) at 6 and 14 
weeks and bivalent OPV (bOPV) at 6, 10, and 14 weeks.  
 
Infants in Bangladesh are given OPV in the form of bOPV, which protects infants against types 1 and 3. 
There is also OPV that protects against just one type of poliovirus. These are called monovalent OPV, or 
mOPV. Monovalent OPV (mOPV) protects infants against type 2 but is only used in very specific 
situations and not part of routine immunization. After polio disease has been eradicated, the expectation 
is that OPV will no longer be given to infants but will still be used for outbreak response. A new vaccine 
that works to protect infants from type 2, called novel OPV, or nOPV2 will likely replace mOPV2 as the 
vaccine used in places with type 2 outbreaks. But we do not know if it is better to give nOPV2 with bOPV 
to help protect children against all three types of poliovirus, or if giving nOPV2 and bOPV together affects 
how well they protect children against polioviruses.  
 
Why is your infant invited to participate in the study as a vaccine recipient?  
We are talking to you about this study because you have an infant that will need to receive polio 
vaccination to be protected against polio disease.  
 
Why is the infant’s older sibling invited to participate in the study?  
For this study, stool samples will be collected from one sibling of each infant who receives the vaccine. 
This will help us understand how polio vaccine spreads from a child who receives vaccine to other 
children in the household.  
 
Methods and procedures of the study 
If you decide to participate, you will need to bring your infant, the one who will receive the vaccine, to 
the study clinic a total of four times over a period of 12 weeks, including today’s visit. Today’s visit 
requires both the vaccine recipient and the sibling to be present, but for the additional visits you will 
only need to bring the vaccine recipient. The additional three clinic visits will be when the vaccine 

 
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recipient is 10, 14, and 18 weeks of age. Each visit will take about 60 minutes. During today’s visit, a 
doctor will ask questions about your children’s health, their vaccinations, and will examine them. For 
the vaccine recipient, the doctor will also administer study vaccines, collect samples, administer routine 
childhood vaccines, observe for any reactions to the vaccines, and schedule the next visit; the same 
activities will take place at the other three study visits. Prior to leaving, the study staff will tell you what 
to do if the vaccine recipient becomes ill, even if the illness is not related to the study vaccines. In 
addition, you will be visited or contacted by phone within 2 days of your infant receiving the first study 
vaccination. The purpose will be for added safety, to check if your infant is experiencing any problems 
or symptoms.  
 
There are three different groups in this study. A computer will choose your infant’s study group. The 
doctor will not know your infant’s study group until the computer does the selection. The doctor will tell 
you the group assignment. The study vaccines are novel OPV2 (nOPV2) and bOPV. Vaccine recipients 
will receive either nOPV2 only, nOPV2 plus bOPV, or bOPV only at 6, 10, and 14 weeks of age. The 
sibling of the vaccine recipient will not receive any study vaccines. 
 
The doctor or nurse will collect a sample of blood (1 ml or less than a quarter teaspoon) each time the 
vaccine recipient comes for a study clinic visit. You will also be asked to collect small stool samples (8 
gm or about the size of an adult thumb) from the vaccine recipient and the enrolled sibling. This will tell 
us how the vaccine works in the intestines and how it can spread between children in the same household.  
 
For the stool collections, study staff will make visits to your home at 2-3 timepoints, when the vaccine 
recipient is 7, 8, or 10 weeks of age. The timing of the stool collections will depend on the vaccine 
recipient’s study group. There will be no stool collection from your children after 10 weeks of age. For 
each stool collection, there will be two or more visits to your household. At the first visit, study staff will 
give you kits to collect stool from your two children. During the next home visit(s), study staff will pick 
up the stool kits.  
 
Once study activities are finished, the doctor or nurse will advise you to vaccinate the vaccine recipient 
with two doses of fIPV. The vaccine recipient may also be advised to receive three doses of bOPV, 
depending on which group he/she is assigned. These additional doses will make sure that your infant has 
protection in the blood and intestines against all three types of poliovirus, like other children in 
Bangladesh. The additional doses will not harm your infant. The doctor or nurse will provide the vaccines 
and will help you schedule the appointments to come to the clinic.  
  
Risk and benefits 
There are benefits from participating in the study. At each visit, a doctor will examine the vaccine 
recipient and administer polio vaccines at no charge. Other vaccines against childhood diseases will also 
be provided to the vaccine recipient during the visits at no cost. These include the pentavalent vaccine 
that protects against diphtheria, tetanus, whooping cough, hepatitis B and Haemophilus influenzae B, and 
the pneumococcal conjugate vaccine that protects against Streptococcus pneumoniae. This study will also 
help the global community because the results will be used to guide how outbreak response activities are 
conducted around the world. 
 
For the vaccine recipient, there are risks to participating in the study. bOPV contains very small amounts 
of antibiotics and an allergic reaction to the antibiotics is possible, but very unlikely. nOPV2 does not 
contain antibiotics. During the study, the vaccine recipient may receive a polio vaccine that is different 
from other children in Bangladesh. bOPV protects against types 1 and 3 and is given to all infants in 

 
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Bangladesh. nOPV2 only protects against type 2 and is not a part of routine immunization. Since nOPV2 
is a new vaccine, we have limited data on its safety, although all the data we have from studies suggests 
that it is safe. We also do not know the duration or extent to which nOPV2 protects against type 2. To 
ensure that all vaccine recipients in the study are protected against all types of poliovirus, after the study 
is over your infant will be given two doses of fIPV, which protects against all three types and is given to 
all infants in Bangladesh through EPI. Your infant will also be given three doses of bOPV based on their 
study group. This means the risk of your infant becoming sick with any of the poliovirus types is very 
low.  
 
Although polio-like disease is a risk with OPV, only one in 1-2 million children develop this kind of 
illness. There is a risk of polio-like disease for the vaccine recipient. We do not know the exact risk of 
polio-like disease in children who receive nOPV2 or bOPV, although we believe it is similar and very 
low. There may also be risk of polio-like disease for others in close contact with the vaccine recipient, 
including the older sibling, but we believe this risk to be very low.  
 
Drawing blood may cause discomfort or pain, and localized bruising, although pressure on the site may 
prevent or reduce bruising. Although rare, it is possible to get an infection at the site of blood draw or 
injection. Trained doctors and nurses will use sterile needles and clean materials to collect blood and 
administer vaccines in order to lower the chance of infection and bruising.   
 
In addition to the expected reactions, there is a chance that the vaccine recipient may experience an 
unexpected reaction. If the vaccine recipient suffers any illness or injury from participating in the study, 
free medical care and treatment will be provided in the appropriate hospital. icddr,b will pay the cost of 
the medical care. In addition, while enrolled in the study, both of your children (the vaccine recipient and 
older sibling) will receive free medical care for any illnesses or injuries that occur, including those that 
are unrelated to participating in the study. If either of your children need medical assistance during the 
study, you may come to icddr,b’s health facilities. A study doctor will examine your child and provide 
medical care according to his/her judgment and local standards. 
 
Privacy, anonymity and confidentiality 
Only a few, select study staff will have access to you and your children’s personal information, such as 
name and address. They will need this information to contact you and to follow up with your children. 
All other study staff will use coded numbers assigned to each child. This includes staff testing the blood 
and stool samples. If the results of the study are published, you and your children will not be identified 
by name.  
 
If the vaccine recipient experiences a severe reaction to the vaccine, his/her records may need to be 
reviewed by other individuals to make sure the study is safe. These people may include the lead study 
investigator, the safety monitor, or a member of the regulatory agency in Bangladesh. These individuals 
will not share names or other personal information with anyone not involved in the review. 
 
Future use of information 
Information collected in this study may be of use to other researchers, vaccine manufacturers and/or 
regulatory authorities (national and international). Upon request to icddr,b, information may be shared 
with these groups in the future. However, you and your children’s names and other personal information 
will not be shared, coded numbers will be shared. This will maintain the same level of privacy, anonymity 
and confidentiality of information that you and your child will have during the study.  
 
Once testing on blood and stool samples are completed and the results are analysed, it may be necessary 
to carry out further testing on the samples. Stored samples will be used for further testing, no additional 

 
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sample collection will be done. If you consent to allow your children to participate in the study, you are 
also consenting to this future testing, if necessary. 
 
Some of your children’s blood and stool samples will be stored for five years in the icddr,b laboratory. 
They will be stored because they contain valuable information about protection in children that may be 
useful for another polio study or other vaccine-preventable disease study. Investigators of future studies 
will need to obtain permission from icddr,b’s research and ethical review committees before they can use 
any stored samples. Genetic testing will not be performed. 
 
 
Right not to participate and withdraw 
Your consent for your children to participate in this study is voluntary. Your children do not have to take 
part in this study. Your children may stop taking part at any time during the study. If your children do not 
participate in the study, they will still have access to vaccines, medical services and other benefits 
provided by the Government of Bangladesh. 
 
Study staff may take a vaccine recipient or sibling out of the study early if: 
• The doctor thinks that the vaccine recipient is too sick to receive one of the study vaccine doses 
• He/she receives a polio vaccine from another source before or during the study 
• You withdraw your consent for the vaccine recipient to be in the study 
 
Principle of compensation  
No monetary compensation will be provided for participating in this study. Money will be provided to 
pay for the cost of transportation to and from the study clinic. Vaccine recipients and siblings will be 
provided free medical care for illnesses and injuries while enrolled in the study.  
 
Answering your questions/ Contact persons 
If you have any questions or concerns about the study or information form, you may speak to:  
 
Name of local principal investigator: Dr. K. Zaman,  
Phone: 01713047100 
 
If you have questions about your child’s rights as a vaccine recipient in this research study or think that 
your child may have been harmed, you may contact the IRB Secretariat of the Ethics Review 
Committee for Research in Human Subjects, icddr,b: 
 
Name: M A Salam Khan  
 
 
 
Phone: 9886498  
 
 
 

 
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Vaccine recipient and Sibling  Informed Consent Form 
 
Protocol No. PR-20060 
Version No. 1.06 
Date: 03-12-2020 
                                 
 
 
 
 
 
Protocol Title: Immunogenicity of novel monovalent oral poliovirus vaccine type 2 (nOPV2) with and 
without bivalent OPV 
Investigator’s name: Dr. K. Zaman  
Organization: International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b) 
 
I have read the  informed consent information sheet, have had the opportunity 
to ask questions, discuss the study, and have received satisfactory answers.    
Yes 
    No 
 
I understand that I am free to withdraw my children from the study without 
giving any reason. 
Yes 
    No 
 
I agree to the collection of up to 1 ml of blood (vaccine recipient) and 8 gm of 
stool (vaccine recipient and older sibling) during each scheduled sample 
collection.  
Yes 
    No 
 
I agree that unidentified blood/stool samples can be sent to CDC for analysis. 
Yes 
    No 
 
I agree for icddr,b to store blood /stool samples up to five years for future use. 
Yes 
    No 
 
I understand that the information I give is confidential. 
Yes 
    No 
 
I agree to my non-identifiable data being used for future, ethically approved 
studies. 
Yes 
    No 
 
I agree to being contacted in the future if there are any studies related to this 
research.  I understand that relevant sections of my children’s medical notes and 
data collected during the study may be looked at by individuals from the sponsor 
organization and by regulatory authorities, where it is relevant to my children 
taking part in this research. I give my permission for those individuals to have 
access to my children’s records.    
Yes 
    No 
 
I give my consent for my infant (Name: ______________) to take part in the 
study and receive polio vaccines. 
Yes 
    No 
 
I give my consent for his/her sibling (Name: _______________) to also take 
part in this study. 
Yes 
    No 
 
 
 
 

 
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If you agree to our proposal of enrolling your infant (Name:_______________) in our study to receive 
polio vaccines, please indicate that by putting your signature or your left thumb impression at the specified 
space below. 
 
Thank you for your cooperation. 
 
 
_______________________________________ 
 
         ____________________ 
Signature or left thumb impression of 
 
  
 
 
Date 
Parent 
 
 
If you also agree to have his/her sibling (Name:_______________) to participate in this study, please 
indicate that by putting your signature or your left thumb impression at the specified space below. 
 
 
_______________________________________ 
 
         ____________________ 
Signature or left thumb impression of 
 
  
 
 
Date 
Parent 
 
 
 
_______________________________________ 
 
         ____________________ 
Signature of the witness 
 
 
 
 
 
 
Date 
 
 
 
_______________________________________ 
 
        ___________________ 
 Signature of the PI or his/her representative  
  
 
            Date 
 
 
Answering your questions/ Contact persons 
If you have any questions or concerns about the study, information form, or consent form, you may 
speak to:  
 
Name of local principal investigator: Dr. K. Zaman,  
Phone: 01713047100 
 
If you have questions about your child’s rights as a vaccine recipient in this research study or think that 
your child may have been harmed, you may contact the IRB Secretariat of the Ethics Review 
Committee for Research in Human Subjects, icddr,b: 
 
Name: M A Salam Khan  
 
 
 
Phone: 9886498  
 
 
 
 
 
 
 
 

 
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Appendix A: Study Forms 
 
• Screening Form 
• Clinic Visit at 6 weeks of Age – Vaccine Recipient 
• Clinic Visit at 6 weeks of Age – Sibling 
• Phone Call or Home Visit 24–48 hours after 6-week Vaccination 
• Household Visit at 7 weeks of Age 
• Household Visit at 8 weeks of Age 
• Household Visit at 10 weeks of Age 
• Clinic Visit at 10 weeks of Age  
• Clinic Visit at 14 weeks of Age  
• Clinic Visit at 18 weeks of Age 
• Adverse Event Form 
• Discontinuation Tracking Form – Vaccine Recipient 
• Discontinuation Tracking Form – Sibling 
• Protocol Deviation Tracking Form 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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nOPV2 and bOPV Study 
 
 
 
    
 
 
PID 
 
Screening Form 
 
Date of interview (dd/MMM/yy): 
                                                    
Vaccine recipient’s name: _____________________________________________ 
 
Date of birth (dd/MMM/yy):                                                                               Sex:  F  /  M    
Demographics  
Father’s education (circle): No formal school  /  Primary  /  Middle   /  High school   /  Graduate  
Mother’s education (circle): No formal school  /  Primary  /  Middle   /  High school   /  Graduate  
Eligibility criteria for vaccine recipient 
1. Child’s residence within the study area? 
 
 
 
 
  
Yes  /   No  
2. Child NOT planning to travel/move in the next 3 months that would prevent them for coming for study visits? 
 
 
 
 
 
 
 
 
 
 
 
Yes  /   No  
3. Child born in single birth?  
 
 
 
 
 
 
Yes  /   No  
4. Child born after 37 weeks of pregnancy? 
 
 
 
 
 
Yes  /   No 
5. At least one sibling <10 years living in the household? 
 
 
 
Yes  /   No 
Is this child eligible for participation in the study as a vaccine recipient?   Yes   /   No    
➢ Child is ELIGIBLE if all answers to questions 1-5 are YES, proceed to question 6. 
➢ Child is NOT ELIGIBLE if any of the answers to questions 1-5 is NO. If the child is NOT eligible, STOP.  
6. How many siblings does the vaccine recipient have that are <10 years of age?       ________ 
➢ For each sibling <10 years of age, complete the eligibility questions below. Then, ask the mother to bring the 
youngest eligible sibling to the Clinic Visit at 6 Weeks of Age, along with the sibling’s vaccination records.  
Eligibility criteria for sibling 
Siblings’ name: _____________________________________________ 
 
Date of birth (dd/MMM/yy):                                                                                
 
Sex:  F  /  M    
1. Child will be <10 years of age at the time of enrollment (when the vaccine recipient is 42-48 days of age)? Yes /  No 
2. Child resides in the same household as the eligible vaccine recipient? 
 
 
Yes  /   No  
3. Child will be available to participate in study activities for the next month ?   
 
Yes  /   No  
4. Child has NOT participated in a previous poliovirus vaccine clinical trial. 
 
 
Yes  /   No  

 
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  PID  
 
Screening Form (page for additional siblings) 
Eligibility criteria for sibling 
Siblings’ name: _____________________________________________ 
 
Date of birth (dd/MMM/yy):                                                                                
 
Sex:  F  /  M    
1. Child will be <10 years of age at the time of enrollment (when the vaccine recipient is 42-48 days of age)? Yes /  No 
2. Child resides in the same household as the eligible vaccine recipient? 
 
 
Yes  /   No  
3. Child will be available to participate in study activities for the next month ?   
 
Yes  /   No  
4. Child has NOT participated in a previous poliovirus vaccine clinical trial. 
 
 
Yes  /   No  
 
Eligibility criteria for sibling  
Siblings’ name: _____________________________________________ 
 
Date of birth (dd/MMM/yy):                                                                                
 
Sex:  F  /  M    
1. Child will be <10 years of age at the time of enrollment (when the vaccine recipient is 42-48 days of age)? Yes /  No 
2. Child resides in the same household as the eligible vaccine recipient? 
 
 
Yes  /   No  
3. Child will be available to participate in study activities for the next month ?   
 
Yes  /   No  
4. Child has NOT participated in a previous poliovirus vaccine clinical trial. 
 
 
Yes  /   No  
 
Eligibility criteria for sibling  
Siblings’ name: _____________________________________________ 
 
Date of birth (dd/MMM/yy):                                                                                
 
Sex:  F  /  M    
1. Child will be <10 years of age at the time of enrollment (when the vaccine recipient is 42-48 days of age)? Yes /  No 
2. Child resides in the same household as the eligible vaccine recipient? 
 
 
Yes  /   No  
3. Child will be available to participate in study activities for the next month ?   
 
Yes  /   No  
4. Child has NOT participated in a previous poliovirus vaccine clinical trial. 
 
 
Yes  /   No  
 
 

 
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   PID  
 
Screening Form 
 
 Is one or more of the children eligible to be a sibling participant?   Yes   /   No    
➢ Child is ELIGIBLE if all answers to sibling eligibility questions 1-4 are YES. If multiple children are eligible, 
select the youngest eligible sibling for participation in the trial. Explain what the child’s participation in the 
study will involve and provide the information sheet. 
➢ Child is NOT ELIGIBLE if any of the answers to sibling eligibility questions 1-4 is NO. If NONE of the 
children are eligible to be sibling participants, STOP. 
 
Are parents interested in their children participating in the study?   Yes   /   No    
➢ If parents ARE NOT interested, ask them the following.  
Why are you not interested in having your children participate in the study? (Circle all that apply) 
 
Study purpose unclear  / Busy-No time  /  Other (specify):_________________________________ 
 
Name of father:  ________________________________Name of mother:  
 
________ 
 
 
Address:  
 
 
 
Section _______ 
Upzila:___ 
 
District:   
 
 
Cell phone:   
____  Scheduled date for first study visit (dd/MMM/yy): 
Name and signature of study staff:   
 
 
 
 
 
____________      
  
 
 
 
 

 
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     PID 
Clinic Visit at 6 weeks of Age – Vaccine Recipient 
 
Date of visit (dd/MMM/yy):  
                                                   
Date of birth (dd/MMM/yy): 
 
Sex:  F  /  M    
Demographics  
Father’s education (circle): No formal school  /  Primary  /  Middle   /  High school   /  Graduate  
Mother’s education (circle): No formal school  /  Primary  /  Middle   /  High school   /  Graduate  
Eligibility criteria for vaccine recipient  
1. Child’s residence within the study area? 
 
 
 
 
 
 
Yes  /   No  
2. Child 6 weeks of age?  
 
 
 
 
 
 
 
 
Yes  /   No  
3. Child and family NOT planning to travel / move in the next 3 months that would prevent them from coming for study 
follow-up visits?  
 
 
 
 
 
 
 
 
 
Yes  /   No  
4. Child born in single birth?  
 
 
 
 
 
 
 
Yes  /   No  
5. Child born after 37 weeks of pregnancy? 
 
 
 
 
 
 
Yes  /   No 
6. No diagnosis or suspicion of immunodeficiency in the child or family member? 
 
Yes  /   No 
7. Child has NO known coagulation disorders that contraindicates venipuncture or injection? 
Yes  /  No 
 
8. Child has NOT received OPV or IPV before? 
 
 
 
 
 
Yes  /  No 
9. Child does not have a medical condition that contraindicates bOPV or nOPV2 administration?     Yes  /  No 
 
If no, specify _________________________________________ 
10. Child has at least one sibling < 10 years living in the household?   
 
 
Yes  /  No 
Is this child eligible for participation in the study as a vaccine recipient?    Yes   /   No    
 
➢ Child is ELIGIBLE if all answers to questions 1-10 are YES. If the child is eligible, proceed to the next 
section.  
➢ Child is NOT ELIGIBLE if any of the answers to questions 1-10 is NO. If the child is NOT eligible, STOP.  
Do the parents give consent for their child to participate in the study as a vaccine recipient?    Yes   /   No    
➢ If parents DO NOT consent, ask them the following. When finished, thank them for their time and STOP.  
Why are you not interested in having your child participate in the study? (Circle all that apply) Study purpose unclear  
/  Vaccine concerns  /  Reluctant to blood draw  /  More than two vaccine injections at first visit / Busy-No time  /  
Other (specify):_________________________________ 
➢ If parents DO consent, continue with the study procedures.    

 
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         PID  
 
Clinical History 
1. Is the child currently breastfed?   No breastfeeding / Partial breastfeeding   / Exclusive breastfeeding 
2. Has the child received BCG?   Yes  /  No     
2a. If yes, date of vaccination (dd/MMM/yy):   
3. Has the child received any other vaccine to date?   Yes  /  No     
3a. If yes, specify: ______________________________________________________________        
  
3b. Date of vaccination (dd/MMM/yy)   
4. Has any other child in the household received OPV since the infant was born?   
Yes / No    
4a. If yes, specify:   
_____________________________________________ 
________ 
5. Has the child had diarrhea in the last 24 hours (≥ 3 loose stools in 24 hours)?   Yes  /  No 
5a. If yes, specify:  duration of diarrhea      
    days 
6. Is the child on any immunosuppressive medications? Yes / No   6a. If yes, specify ______________  
Physical Exam 
7. Temperature:              .       °Celsius  8. Length #1:                    .       cm 10. Weight #1:             .        kg    
              
 
 
 
     9. Length #2:                    .       cm 11.Weight #2:             .        kg 
Blood Collection 
12. Blood sample collected?   Yes / No 
12a. If not, why? __________________________________ 
13. Sufficient quantity (1 ml)?  Yes / No 
13a. If insufficient, why? ____________________________ 
14. Time of collection (hh:mm):            :  
Study Group Allocation and Vaccine Administration 
15. Specify which study group the child was allocated (select the answer):  
A: nOPV2 @ 6, 10, and 14 weeks 
B: nOPV2 + bOPV @ 6, 10, and 14 weeks 
C: bOPV @ 6, 10, and 14 weeks  
 
 
 
 
 
 
 
Clinic Visit at 6 weeks of Age – Vaccine Recipient 

 
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            PID  
 
 
Vaccine Administration  
If Study Arm A or B 
16. nOPV2 administered?          Yes / No   16a. If not, reason: ____________________________ 
If administered: 16b. Time of vaccine administration (hh:mm):                 :  
16c. Expiration (mmm/yy): 
 
If Study Arm B or C 
17. bOPV administered?          Yes / No   17a. If not, reason: __________________________________ 
If administered: 17b. Time of vaccine administration (hh:mm):                 :  
17c. Expiration (mmm/yy): 
 
18. BCG administered?   Yes  /  No     18a. If no and infant has yet to receive BCG, provide reason not  
given during today’s visit: ______________ 
 
 
If administered:  
18b. Site of administration:    Right arm   /   Left arm  
 
19. Pentavalent administered?   Yes  /  No     19a. If not, reason:  ______________ 
 
 
__ 
If administered:  
19b. Dose:    Penta1  /  Penta2  /  Penta3        
19c. Site of administration:    Right thigh   /   Left thigh  
20. PCV administered?    Yes  /  No 
    20a. If not, reason:   
________ 
 
________ 
If administered:  
20b. Dose:    PCV1  /  PCV2  /  PCV3        
20c. Site of administration:   Right thigh   /   Left thigh  
 
Observe for 30 minutes after vaccine administration:   
21. Any symptoms?  Yes  /  No 
21a. If yes, specify (complete Adverse Event Form, if required): ____________________________ 
________________________________________________________________________________ 
  
Schedule next visit date (dd/MMM/yy):   
Name and signature of medical officer:   
 
 
 
 
________________  
 
 
Clinic Visit at 6 weeks of Age – Vaccine Recipient 

 
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               PID  
 
Clinical History 
 
Date of birth (dd/MMM/yy): 
 
 
Current age in years: 
Sex:  F  /  M    
 
Eligibility criteria for sibling 
1. Child is <10 years of age   
 
 
 
 
 
 
 
Yes  /   No 
2. Child resides in the same household as the study vaccine recipient?  
 
 
Yes  /   No  
3. Child will be available to participate in study activities for the next month?   
 
Yes  /   No  
4. Child has NOT participated in a previous poliovirus vaccine clinical trial. 
 
 
Yes  /   No  
Is this child eligible to be a sibling in the study?   Yes   /   No    
 
➢ Child is ELIGIBLE if all answers to questions 1-4 are YES. If the child is eligible, explain what the child’s 
participation in the study will involve and provide the information sheet. 
➢ Child is NOT ELIGIBLE if any of the answers to questions 1-4 is NO. If the child is NOT eligible, STOP. The 
sibling and vaccine recipient are not eligible to continue with study enrollment. Enrollment cannot continue 
for the vaccine recipient unless an eligible sibling is brought to the clinic to be enrolled on the same day.    
Do the parents give consent for their child to participate in the study?    Yes   /   No    
➢ If parents DO NOT consent, ask them the following. When finished, thank them for their time and STOP.  
Why are you not interested in having your child participate in the study? (Circle all that apply) Study purpose unclear  
Busy-No time  /  Other (specify):_________________________________ 
➢ If parents DO consent, continue with the study procedures.    
Polio vaccination history: 
1. Is the vaccination card present?    Yes / No 
Note: If present, answer questions below based on the vaccination card.  
2. Has the child ever received bOPV?    Yes  /  No     
2a. If yes, how many doses of bOPV?    ______________ 
2b. Date of most recent bOPV vaccination (dd/MMM/yy):    
 
 
Clinic Visit at 6 weeks of Age – Sibling  

 
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Clinical History 
 
3. Has the child ever received tOPV?    Yes  /  No     
3a. If yes, how many doses of tOPV?    ______________ 
3b. Date of most recent tOPV vaccination (dd/MMM/yy):    
4. Has the child ever received fIPV?    Yes  /  No     
4a. If yes, how many doses of fIPV?    ______________ 
4b. Date of most recent fIPV vaccination (dd/MMM/yy):    
5. Has the child ever received IPV?    Yes  /  No     
5a. If yes, how many doses of IPV?    ______________ 
5b. Date of most recent IPV vaccination (dd/MMM/yy):    
 
Name and signature of medical officer:   
 
 
 
 
________________ 
 
 
 
 
Clinic Visit at 6 weeks of Age – Sibling  

 
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             PID 
Phone Call or Home Visit 24–48 hours after 6-week vaccination 
To be completed for vaccine recipients in all Arms 
   Date of call/visit (dd/MMM/yy):  
 
 
Time of call/visit (hh:mm): 
 
Adverse events 
Since the vaccine recipient was at the clinic for the “Clinic Visit at 6 weeks”, have they had any of the following 
symptoms? 
1. Fever: 
 No  
  Yes      If yes, please describe ______________________________________ 
If yes, and thermometer available: 
 Grade 1: 37.5° C- 38.0 ° C  
 Grade 2: 38.1° C- 39.0 ° C
 
 
 
 
    
 Grade 3: >39.0 ° C 
 
2. Vomiting:  
 No  
  Yes      If yes,  
 Grade 1: 1 episode/24 hours  
 Grade 2: 2-5 episodes/24 hours
  
 
 
 
 
 Grade 3:  ≥ 6 episodes/24 hours 
 
3. Abnormal crying:  
 No  
  Yes      If yes,  
 Grade 1: < 1hour     
 Grade 2: 1-3 hours 
 
 Grade 3: >3 hours 
 
 
4. Drowsiness:  
 No  
  Yes      If yes, 
 Grade 1: Sleepier than usual or less interested in surroundings 
 Grade 2: Not interested in surroundings or did not wake up for a feed 
 Grade 3: Sleeping most of the time or difficult to wake up 
 
5. Poor feeding:  
 No  
  Yes      If yes, 
 Grade 1: Eating less than normal 
 Grade 2: Missed 1 or 2 feeds completely  
 Grade 3: Refuses ≥3 feeds or refuses most feeds 
 
6. Irritability:  
 No  
  Yes      If yes, 
 Grade 1: Easily consolable 

 
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 Grade 2: Requiring increased attention 
 Grade 3: Inconsolable 
7. Other Adverse Events (24–48 hours after 6-week vaccination)    
 No  
  Yes       
If yes, specify: ______________________________________ 
If yes:   
 Grade 1 (Mild): An AE which is easily tolerated by the participants, causing      
minimal discomfort and not interfering with everyday activities.  
 
 Grade 2 (Moderate): An AE which is sufficiently discomforting to interfere with 
normal everyday activities.     
 
 Grade 3 (Severe): An AE which prevents normal, everyday activities. 
If parent responds Yes to any of the above, fill out an AE form.  
Name of medical officer:_____________________________ Signature: ____________________________ 
 
 

 
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Household Visit at 7 weeks of Age 
To be completed for participants in Arm A only 
Home Visit #1:  To be completed on the day the stool kits are delivered to the parents. 
Date stool kits delivered (dd/mm/yy): 
1. Were there any problems in giving the two stool kits to the parents?  Yes  /  No    
1a. If yes, explain ________________________________________________________ 
 
Name and signature of study staff:   
 
 
 
 
 
 
 
 
 
Home Visit #2: To be completed on the day the stool samples are picked up. 
For vaccine recipient: 
1. Stool sample collected?   Yes / No      
1a. If no stool obtained, why not? ____________________ 
2. Date of stool collection (dd/mm/yyyy):            
3. Time of collection (hh:mm):              : 
 
4. Sufficient quantity (8 grams)?  Yes / No 
 4a. If insufficient, specify reasons: ___________________ 
 
For sibling: 
5. Stool sample collected?   Yes / No    5a. If no stool obtained, why not? ____________________ 
6. Date of stool collection (dd/mm/yyyy):            
7. Time of collection (hh:mm):              : 
 
8. Sufficient quantity (8 grams)?  Yes / No 
 8a. If insufficient, specify reasons: ___________________ 
 
Name and signature of study staff:   
 
 
 
 
 
 
 
 
 
Data Entry: To be completed when data entered into RedCap 
Date of data entry in RedCap (dd/mm/yyyy): 
 
Name and signature of person entering data:    
 
 
 
 
 
 
 
 
 

 
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Household Visit at 8 weeks of Age 
To be completed for vaccine recipients in all Arms 
Home Visit #1:  To be completed on the day the stool kits are delivered to the parents. 
Date stool kits delivered (dd/mm/yy): 
1. Were there any problems in giving the two stool kits to the parents?  Yes  /  No    
1a. If yes, explain ________________________________________________________ 
 
Name and signature of study staff:   
 
 
 
 
 
 
 
 
 
Home Visit #2: To be completed on the day the stool samples are picked up. 
For study vaccine recipient: 
1. Stool sample collected?   Yes / No      
1a. If no stool obtained, why not? ____________________ 
2. Date of stool collection (dd/mm/yyyy):            
3. Time of collection (hh:mm):              : 
 
4. Sufficient quantity (8 grams)?  Yes / No 
 4a. If insufficient, specify reasons: ___________________ 
 
For sibling of study vaccine recipient: 
5. Stool sample collected?   Yes / No    5a. If no stool obtained, why not? ____________________ 
6. Date of stool collection (dd/mm/yyyy):            
7. Time of collection (hh:mm):              : 
 
8. Sufficient quantity (8 grams)?  Yes / No 
 8a. If insufficient, specify reasons: ___________________ 
 
Name and signature of study staff:   
 
 
 
 
 
 
 
 
 
Data Entry: To be completed when data entered into RedCap 
Date of data entry in RedCap (dd/mm/yyyy): 
 
Name and signature of person entering data:    
 
 
 
 
 
 
 
 
 

 
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Household Visit at 10 weeks of Age 
To be completed for vaccine recipients in all Arms 
Home Visit #1:  To be completed on the day the stool kits are delivered to the parents. 
Date stool kits delivered (dd/mm/yy): 
1. Were there any problems in giving the two stool kits to the parents?  Yes  /  No    
1a. If yes, explain ________________________________________________________ 
 
Name and signature of study staff:   
 
 
 
 
 
 
 
 
 
Home Visit #2: To be completed on the day the stool samples are picked up. 
For study vaccine recipient: 
1. Stool sample collected?   Yes / No      
1a. If no stool obtained, why not? ____________________ 
2. Date of stool collection (dd/mm/yyyy):            
3. Time of collection (hh:mm):              : 
 
4. Sufficient quantity (8 grams)?  Yes / No 
 4a. If insufficient, specify reasons: ___________________ 
 
For sibling of study vaccine recipient: 
5. Stool sample collected?   Yes / No    5a. If no stool obtained, why not? ____________________ 
6. Date of stool collection (dd/mm/yyyy):            
7. Time of collection (hh:mm):              : 
 
8. Sufficient quantity (8 grams)?  Yes / No 
 8a. If insufficient, specify reasons: ___________________ 
 
Name and signature of study staff:   
 
 
 
 
 
 
 
 
 
Data Entry: To be completed when data entered into RedCap 
Date of data entry in RedCap (dd/mm/yyyy): 
 
Name and signature of person entering data:    
 
 
 
 
 
 
 
 
 

 
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Clinic Visit at 10 weeks of Age 
Date of visit (dd/mm/yy):  
 
Clinical History 
 
1. Is the child currently breastfed?  No breastfeeding / Partial breastfeeding / Exclusive breastfeeding 
2. Has the child received any vaccine since last visit?   Yes  /  No   2a. If yes, specify:  _____________   
2b. Date of vaccination (dd/mm/yy):    
3. Has any other child in the household received OPV since the last clinic visit ?   
Yes / No   3a. If yes, 
specify: ______________ 
4. Has the child had diarrhea in the last 24 hours (> 3 loose stools in 24 hours)?   Yes  /  No 
4a. If yes, specify: duration of diarrhea in days:               
(If the child currently has fever >38.3°C, blood in stools, vomiting or signs of dehydration, do not 
continue with study procedures. Provide advice and referral as required.)      
5. Has the child been sick since the last study visit?  Yes / No   5a. If yes, specify: __________________ 
6. Is the child on any immunosuppressive medications?   Yes  /  No   6a. If yes, specify ___________ 
Physical Exam 
7. Temperature:              .       °Celsius   8. Length #1:                    .       cm    10. Weight #1:             .      kg                     
              
 
 
 
       9. Length #2:                 .        cm   11. Weight #2:             .      kg 
Blood Collection 
12. Blood sample collected?   Yes / No 
12a. If not, why? __________________________________ 
13. Sufficient quantity (1 ml)?  Yes / No 
13a. If insufficient, why? ____________________________ 
14. Time of collection (hh:mm):            :  
 
Vaccine Administration  
If Study Arm A or B 
15. nOPV2 administered?          Yes / No   15a. If not, reason: ____________________________ 
If administered: 15b. Time of vaccine administration (hh:mm):                 :  
15c. Expiration (mmm/yy): 
If Study Arm B or C 
16. bOPV administered?          Yes / No   16a. If not, reason: __________________________________ 
If administered: 16b. Time of vaccine administration (hh:mm):                 :  
16c. Expiration (mmm/yy): 

 
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Clinic Visit at 10 weeks of Age (ctd.) 
 
17. Pentavalent administered?   Yes  /  No     17a. If not, reason:  ______________ 
 
 
__ 
If administered:  
17b. Dose:    Penta1  /  Penta2  /  Penta3        
17c. Site of administration:    Right thigh   /   Left thigh  
18. PCV administered?    Yes  /  No 
    18a. If not, reason:   
________ 
 
________ 
If administered:  
18b. Dose:    PCV1  /  PCV2  /  PCV3        
18c. Site of administration:   Right thigh   /   Left thigh  
Observe for 30 minutes after vaccine administration:   
19. Any symptoms?  Yes  /  No 
19a. If yes, specify (complete Adverse Event Form, if required): ____________________________ 
________________________________________________________________________________ 
 
The following questions correspond to sibling of the study vaccine recipient. Note: the sibling does not 
need to be present for this visit.  
20. Has the child received any poliovirus vaccines (bOPV, fIPV) since last visit?   Yes  /  No    
20a. If yes, specify:  
_____________   
20b. Date of vaccination (dd/mm/yy):    
21. Has the child had diarrhea in the last 24 hours (> 3 loose stools in 24 hours)?   Yes  /  No 
21a. If yes, specify: duration of diarrhea in days:               
22. Has the child been sick since the last study visit?  Yes / No   22a. If yes, specify: __________________ 
23. Is the child on any immunosuppressive medications?   Yes  /  No   23a. If yes, specify ___________ 
 
Schedule next visit date (dd/mm/yy):   
 
Name and signature of medical officer:  ______________________________________________ 
 
 
 

 
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PID 
Clinic Visit at 14 weeks of Age 
Date of visit (dd/mm/yy):  
 
Clinical History 
 
1. Is the child currently breastfed?  No breastfeeding / Partial breastfeeding / Exclusive breastfeeding 
2. Has the child received any vaccine since last visit?   Yes  /  No   2a. If yes, specify:  _____________   
2b. Date of vaccination (dd/mm/yy):    
3. Has any other child in the household received OPV since the last clinic visit?   
Yes / No   3a. If yes, 
specify: ______________ 
4. Has the child had diarrhea in the last 24 hours (> 3 loose stools in 24 hours)?   Yes  /  No 
4a. If yes, specify: duration of diarrhea in days:               
(If the child currently has fever >38.3°C, blood in stools, vomiting or signs of dehydration, do not 
continue with study procedures. Provide advice and referral as required.)      
5. Has the child been sick since the last study visit?  Yes / No   5a. If yes, specify: __________________ 
6. Is the child on any immunosuppressive medications?   Yes  /  No   6a. If yes, specify ___________ 
Physical Exam 
7. Temperature:              .       °Celsius   8. Length #1:                    .       cm    10. Weight #1:             .      kg                     
              
 
 
 
       9. Length #2:                 .        cm   11. Weight #2:             .      kg 
 
Blood Collection 
12. Blood sample collected?   Yes / No 
12a. If not, why? __________________________________ 
13. Sufficient quantity (1 ml)?  Yes / No 
13a. If insufficient, why? ____________________________ 
14. Time of collection (hh:mm):            :  
 
Vaccine Administration  
If Study Arm A or B 
15. nOPV2 administered?          Yes / No   15a. If not, reason: ____________________________ 
If administered: 15b. Time of vaccine administration (hh:mm):                 :  
15c. Expiration (mmm/yy): 

 
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Clinic Visit at 14 weeks of Age (ctd.) 
 
If Study Arm B or C 
16. bOPV administered?          Yes / No   16a. If not, reason: __________________________________ 
If administered: 16b. Time of vaccine administration (hh:mm):                 :  
16c. Expiration (mmm/yy): 
 
17. Pentavalent administered?   Yes  /  No     17a. If not, reason:  ______________ 
 
 
__ 
If administered:  
17b. Dose:    Penta1  /  Penta2  /  Penta3        
17c. Site of administration:    Right thigh   /   Left thigh  
 
18. PCV administered?    Yes  /  No 
    18a. If not, reason:   
________ 
 
________ 
If administered:  
18b. Dose:    PCV1  /  PCV2  /  PCV3        
18c. Site of administration:   Right thigh   /   Left thigh  
 
Observe for 30 minutes after vaccine administration:   
19. Any symptoms?  Yes  /  No 
19a. If yes, specify (complete Adverse Event Form, if required): ____________________________ 
________________________________________________________________________________ 
 
Schedule next visit date (dd/mm/yy):   
 
Name and signature of medical officer:  ______________________________________________ 
 
 
 
 

 
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Clinic Visit at 18 weeks of Age 
Date of visit (dd/mm/yy):  
 
Clinical History 
 
1. Is the child currently breastfed?  No breastfeeding / Partial breastfeeding / Exclusive breastfeeding 
2. Has the child received any vaccine since last visit?   Yes  /  No   2a. If yes, specify:  _____________   
2b. Date of vaccination (dd/mm/yy):    
3. Has any other child in the household received OPV since the last clinic visit?   
Yes / No   3a. If yes, 
specify: ______________ 
4. Has the child had diarrhea in the last 24 hours (> 3 loose stools in 24 hours)?   Yes  /  No 
4a. If yes, specify: duration of diarrhea in days:               
(If the child currently has fever >38.3°C, blood in stools, vomiting or signs of dehydration, do not 
continue with study procedures. Provide advice and referral as required.)      
5. Has the child been sick since the last study visit?  Yes / No   5a. If yes, specify: __________________ 
6. Is the child on any immunosuppressive medications?   Yes  /  No   6a. If yes, specify ___________ 
Physical Exam 
7. Temperature:              .       °Celsius   8. Length #1:                    .       cm    10. Weight #1:             .      kg                     
              
 
 
 
       9. Length #2:                 .        cm   11. Weight #2:             .      kg 
 
Blood Collection 
12. Blood sample collected?   Yes / No 
12a. If not, why? __________________________________ 
13. Sufficient quantity (1 ml)?  Yes / No 
13a. If insufficient, why? ____________________________ 
14. Time of collection (hh:mm):            :  
 
 
Schedule next visit date to begin routine polio vaccination schedule (dd/mm/yy):   
 
Name and signature of medical officer:  ______________________________________________ 
 
 
 
 
 
 

 
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Adverse Event Form 
Date of report (dd/mm/yy): 
Report Status:         Initial Report             Follow-up Report             Final report 
Initials of parent providing information:   
Vaccine recipient date of birth (dd/mm/yy): 
Description of the event 
Is this a solicited (from the 24–48 hours after 6-week vaccination) or unsolicited AE?   Solicited  /  Unsolicited 
Date of onset (dd/mm/yy): 
 
 
 
                  Time of onset (hh:mm):              :     
Describe briefly relevant clinical symptoms and signs:   
 
 
 
 
 
 
 
Results of diagnostic tests performed:   
 
 
 
 
 
 
 
 
 
Medications prescribed: 
 
 
 
 
 
 
 
 
 
 
 
Diagnosis (circle: Current  /  Final):   
______ 
 
 
 
 
 
 
 
Severity (circle what applies):    Mild   /   Moderate    /   Severe 
Did any of the following occur or are present? (if present, report immediately as a Serious Adverse Event) 
Paralysis, severe disability or incapacity, or substantial  
disruption of the ability to conduct normal functions  
 
Yes / No    
Anaphylaxis  
 
 
 
 
 
 
Yes / No    
Hospitalization or prolongation of existing hospitalization  
Yes / No    
Death  
 
 
 
 
 
 
 
Yes / No    
Life-threatening event  
 
 
 
 
 
Yes / No    
Outcome (circle what applies):   Ongoing  /  Resolved without sequela  /  Resolved with sequela   /  Death  
Date when event resolved or death (dd/mm/yy): 
Did this AE lead to discontinuation from the study or from future vaccinations?    Yes / No    
 
 

 
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Adverse Event Form (cont.) 
Study group allocation: Specify which study group the child was allocated (circle the answer):  
A: nOPV2 @ 6, 10, and 14 weeks 
B: nOPV2 + bOPV @ 6, 10, and 14 weeks 
C: bOPV @ 6, 10, and 14 weeks  
 
Study vaccines administered:    
Clinic visit 
Study vaccines 
administered 
Date (DD/MMM/YY) 
Lot & batch number 
6 weeks of age 
 
___ / _____ / ___ 
 
10 weeks of age 
 
___ / _____ / ___ 
 
14 weeks of age 
 
___ / _____ / ___ 
 
Routine immunization vaccines administered:  
Clinic visit 
Vaccines administered 
At birth 
 
6 weeks of age 
 
10 weeks of age 
 
14 weeks of age 
 
18 weeks of age 
 
At any other age, specify age_______ 
 
 
Signature of study investigator/s:    
 
 
 
 
 
 
 
To be completed by the principal investigator  
Is the event related to the study vaccine?        
       Unrelated 
Possibly related 
Probably related 
Definitely related 
If unrelated to the vaccine, provide a reasonable explanation for the event (e.g., other vaccine, blood 
collection, medical collection, etc.) ______________________________________________________ 
___________________________________________________________________________________ 
___________________________________________________________________________________ 
Name and signature of PI:    
 
 
 
 
  Date 

 
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Discontinuation Tracking Form – Vaccine Recipient  
Date of the report (dd/mm/yy): 
Date of study discontinuation (dd/mm/yy): 
Note: This is the date the vaccine recipient is withdrawn from the study and may not be the same date as the report  
Last clinic visit (circle one):  6 weeks / 10 weeks / 14 weeks  
Date of last study clinic visit (dd/mm/yy):  
Reasons (circle one of the choices listed below): 
 
Discontinuation criteria as mentioned in study protocol: 
1. Parent withdrew consent for participation (i.e., did not want child to participate any further). 
2. The vaccine recipient received polio vaccine outside of the study. (Complete protocol deviation form) 
3. Parent refused to allow collection of blood at 6 weeks of age. (Complete protocol deviation form) 
4. Study staff were unable to collect blood at 6 weeks of age (Complete protocol deviation form) 
5. Since enrolment, the vaccine recipient was diagnosed with a medical condition in which continued 
participation in the study posed a health risk (for example, immunodeficiency or bleeding disorder). 
6. The vaccine recipient began taking immunosuppressive medications. 
7. The PI withdrew or parent refused participation because the vaccine recipient experienced a serious adverse 
event, not death, in which continued participation in the study posed a health risk.  
(Complete Adverse 
Event form) 
8. The PI withdrew or parent refused participation because the vaccine recipient experienced an adverse event, 
not serious AE, in which continued participation in the study posed a health risk.  
(Complete Adverse 
Event form) 
9. Withdrawal by investigator (specify reason): ______________________________________________  
Other reasons why vaccine recipient may not continue in the study: 
10. The vaccine recipient died.  
(Complete Adverse Event form) 
11. The PI withdrew because the parent and vaccine recipient could not be located (i.e., lost to follow-up). 
12. Other (specify reason): _______________________________________________________________ 
__________________________________________________________________________________ 
Name of medical officer:_____________________________ Signature: ____________________________ 
 
 

 
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Discontinuation Tracking Form – Sibling  
Date of the report (dd/mm/yy): 
Date of study discontinuation (dd/mm/yy): 
Note: This is the date the sibling is withdrawn from the study and may not be the same date as the report  
Last study visit (circle one):  6 weeks / 7 weeks  / 8 weeks / 10 weeks  
Date of last study visit (dd/mm/yy):  
Reasons (circle one of the choices listed below): 
 
Discontinuation criteria as mentioned in study protocol: 
1. Vaccine recipient was discontinued from the study. 
2. Parent withdrew consent for sibling’s participation (i.e., did not want child to participate any further). 
3. The sibling received polio vaccine during their participation in the study. (Complete protocol deviation 
form) 
4. Withdrawal by investigator (specify reason): ______________________________________________  
Other reasons why vaccine recipient may not continue in the study: 
5. The sibling died.   
6. The PI withdrew because the parent and sibling could not be located (i.e., lost to follow-up). 
7. Other (specify reason): _______________________________________________________________ 
__________________________________________________________________________________ 
Name of medical officer:_____________________________ Signature: ____________________________ 

 
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Protocol Deviation Tracking Form 
Complete this form for protocol deviations and protocol violations. 
Date of the report (dd/mm/yy): 
Date of protocol deviation (dd/mm/yy): 
Note: This is the date the protocol deviation occurred,  
may not be the same date as the report.  
 
1. Describe the protocol deviation:  ______________________________________________________ _ 
___________________________________________________________________________________ 
___________________________________________________________________________________ 
2. Reason for protocol deviation (circle one answer) 
-    Vaccine recipient illness    
 
 
        
-  Laboratory error 
 
-    Vaccine recipient (or parent) unable to comply              -  Clinic error  
 
-    Parental refusal 
 
 
 
 
-  Investigator/study decision 
 
-    Sibling unable to comply 
 
-    Other,    Specify   _______________________________________________________________ 
3. Deviation category  (circle one answer):  
 
-  Eligibility / enrolment 
 
 
-  Study vaccine type or dosing 
 
-  Protocol procedure / assessment 
 
-  Biological specimen collection or processing 
 
-  Clinic visit schedule 
 
4. Describe steps taken to resolve or avoid recurrence of the deviation: 
 
 
 
 
 
 
 
 
 
 
 
 
 
_____ 
 
 
 
 
 
 
 
 
 
 
 
 
___________ 
5. Did the deviation result in an adverse event?   Yes  /  No    (If Yes, complete an Adverse Event Form) 
6. Did the deviation result in vaccine recipient discontinuation?  Yes  /  No     
(If Yes, complete Vaccine recipient Discontinuation Tracking Form) 
 
Name of medical officer:__________________________ Signature: ____________________________ 
 
 

 
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Appendix B: Microneutralization Test for Polio Antibodies 
 
Prepared by 
Date Adopted 
Supersedes Procedure  
William Weldon 
4/2/2014 
12_10_10 
 
Introduction 
The polio microneutralization assay measures neutralizing antibody titers to poliovirus types 1, 2, and 3 using 96-
well microtiter plates (it is termed “microneutralization” because the original neutralization assay was performed 
using larger volumes on culture tubes). The principle of the test is that the anti-poliovirus antibodies in a serum 
sample will bind to the virus and block infection of susceptible cells. Because poliovirus is cytopathic, virus that is 
not bound by antibody infects and lyses cells. The amount of neutralizing antibody is quantitated as a 50% endpoint 
titer; that is, the dilution of serum that protects 50% of susceptible cells from poliovirus infection and cytopathic 
effect. 
 
The test takes approximately seven days to complete, from the dilution of sera to staining and reading plates, and 
data analysis. Each test serum is run in triplicate and diluted from 1:8 to 1:1024; a single 96-well plate contains 
four test sera (Figure 1). This test may be performed manually, automated, or in a combination of the two 
approaches (Figure 2). For large studies, automation is recommended. 
 
 
 
Figure 1. 96-well plate setup for Polio Serology 
 
For a single run, up to 96 sera, which fit on 72 plates (24 plates per serotype), can be tested. For most purposes, 
the Sabin vaccine strains are used, but other poliovirus strains can be used. Control plates are generated for 
each run and consist of three back titration plates with no antibody added (one for each Sabin virus) and a cell 
control plate (no virus or antibody added, to assess cell viability). At the end of each run, control plates are 
checked for accurate dilution of each Sabin poliovirus (back titration plates) or cell monolayer confluency (cell 
control plate). 
 
If more than seven sera are being tested, the samples must be randomized using a balanced block randomization 
scheme with integrated controls. Included in each run is a control serum designated In-House Reference Serum 
(IHRS), which is pooled from serum samples with high neutralizing antibody titers to each Sabin poliovirus. 

 
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The IHRS is tested in multiple replicates, on multiple plates in each run, to provide a measure of assay 
variability within a run. 
 
 
 
Figure 2. Workflow for automated poliovirus microneutralization assay 
 
Materials and Equipment 
Most of these items may be substituted with equivalent items from other manufacturers/suppliers but alternative 
materials must be validated against an appropriate standard. 
 
Consumables 
• 
T-150 tissue culture flasks (Corning, #430823 ) 
• 
96-well tissue culture, clear, sterile plates (Corning, #3997) 

 
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• 
Low evaporation lids (Corning, #3931) 
• 
96-well tissue culture, clear, sterile, (Corning #3997) 
• 
Plastic wrap (e.g., Saran Wrap) 
• 
Deep-well 96-well microplate, 2.2 ml capacity. (VWR, #40002-014) 
• 
Mat lids for 2.0 mL microplates (VWR, #400002-018) 
• 
Sterile pipettes; 10 ml, 25 ml (Falcon, #357551, #357325) 
• 
Single-channel pipettes:  
o Manual: LTS 20, 200, 1000 (Rainin) 
o Electronic: 10-300, 50-1000 (Biohit) 
• 
12-channel pipettes  
o Electronic: 10-300, 50-1200 (Biohit) 
• 
Pipette tips 
o Rainin: RT-L200F, RT-L1000F, RT-L10F 
o Biohit: 350 μL, 1000 μL, 1200 μL 
• 
WellPro 250 μL sterile pipette tips (ProGroup) 
 
Cells and Media 
• 
HEp-2C cells (ATCC # CCL23) 
• 
Cell culture media 
o Eagle’s Minimum Essential Media (EMEM)(Gibco, #11095-072)  
o Penicillin/streptomycin (Gibco, #15140) 
o Fetal Bovine Serum - Optima (Atlanta Biologicals, #S12450) 
o 0.05% Trypsin-EDTA (Gibco, #25300) 
• 
Nalgene 500 mL, 0.20 μm filter (Nalgene, #450-0020) 
 
Antigens and Control Sera 
• 
In House Reference Sera (developed in-house)(See Note 1.4.4) 
• 
Sabin virus stocks grown in HEp2-C cells 
 
Staining 
• 
Crystal violet stain (0.05% crystal violet, 0.5% Tween-20, 50% ethanol, in H2O) 
 
To prepare crystal violet stock solution: 
2 g 
 
crystal violet (Sigma, C-3386) 
1000 ml 
95% ethanol  
 
Mix together overnight, with stirring, until dissolved; may be stored up to one year at room temperature 
 
To prepare working dilution crystal violet solution: 
 
250 ml  crystal violet stock solution 
 
5 ml 
 
Tween-20 (Fisher Scientific, #BP337-100) 
 
750 ml  
deionized H20  
 
Equipment and Automation 
• 
CO2 water-jacketed incubator (ThermoFisher Model 3110 or equivalent) 
• 
WellPro 4000 CE (ProGroup) 
• 
EL406 plate washer and dispenser (BioTek) 

 
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• 
MicroFlo dispenser (BioTek) 
• 
Evo 100 (Tecan) 
• 
TC20 Automatic Cell Counter (BioRad) 
• 
PerkinElmer Victor X4 Multimode plate reader or equivalent 
 
Protocol 
In the week preceding the test runs: 
 
1. Assign sera randomly to each run using a balanced block randomization scheme (see Note 3). 
2. Use the list generated by the randomization scheme to label plates and organize sera to be tested. 
Each test serum is run in triplicate, so four sera may be run on each plate (Figure 1).  Each plate is 
duplicated two more times, yielding three plates, one for each poliovirus serotype. There will also be 
a back-titration plate for each serotype, and one cell control plate per run. 
3. In this example of a randomized sample list (Table 1), serum sample number 0000000001 will be in 
run number 420, position 4 on plates 5 (PV1), 29 (PV2), and 53 (PV3). The in-house reference serum 
(IHRS) is in position 2 on plates 7 (PV1), 30 (PV2), and 55 (PV3). 
 
RUN
a 
PV1 
Plateb 
PV2 
Plateb 
PV3 
Plateb 
Positionc 
Sample IDd 
420 
5 
28 
53 
1 
0000000001 
420 
5 
28 
53 
2 
0000000002 
420 
5 
28 
53 
3 
0000000006 
420 
5 
28 
53 
4 
0000000009 
420 
6 
29 
54 
1 
0000000011 
420 
6 
29 
54 
2 
0000000030 
420 
6 
29 
54 
3 
0000000025 
420 
6 
29 
54 
4 
0000000041 
420 
7 
30 
55 
1 
0000000013 
420 
7 
30 
55 
2 
IHRS 
420 
7 
30 
55 
3 
0000000038 
420 
7 
30 
55 
4 
0000000051 
 
Table 1. Example of randomized sample list for poliovirus microneutralization assay. 
a each run of 1-96 sera 
b PV1, polio type 1; PV2, polio type 2; PV3, polio type 3 
c see Figure 1 
d unique specimen ID 
 
4. For each run, the IHRS is tested an average of 4-6 times, depending on the number of samples being 
randomized (see Note 4). 
5. The labelling of the assay plates should reflect the setup of the serology run reflected in the 
randomized list. A suggested plate labelling scheme is shown in Figure 3. 
 

 
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Figure 3. Suggested labeling scheme for assay plates. 
 
6. Prepare MEM+ 2% FBS and MEM+10% FBS: 
a. FBS must be inactivated at 56°C for 30 minutes and filtered with a Nalgene 0.2 µm filter 
b. Add 1 mL of streptomycin/penicillin to 1000 mL of EMEM (0.1%) 
c. Add 20 ml (2%) or 100 mL (10%) of fetal bovine serum to 1000 ml bottle of EMEM 
d. MEM+10% FBS is used for maintaining HEp2-C cells and generating the cell suspension. 
e. MEM+2% FBS is used for filling the assay plates, diluting sera, and diluting the virus.  
 
Prior to each test run: 
 
5. 24-48 hours before each run, seed T-150 flasks with 30 ml of HEp2-C cells at 5 x 105 cells/ml.  
a. Approximately 3-4 flasks are needed for one run of 96 sera 
6. Incubate flasks at 37°C, 5% CO2, in a humidified atmosphere for 24 to 48 hours to ensure that cell 
monolayers are confluent the day assay runs are started. 
 
OPTIONAL - One day before test run: 
 
The following steps can be done the day of the run or the day before the addition of the virus. 
 
1. Manually aliquot 100 μl of each serum sample into deep-well polypropylene microplate, sealed with 
a mat lid, and heat inactivated at 56°C in a water bath for 30 minutes. 
2. Store at 4°C until ready to generate the triplicate sample plates (no more than 24 hours). 
3. Prepare IHRS for testing (See Note 4) 
4. Use a multichannel pipette to add 300 μl MEM+2% FBS to 100 μl heat-inactivated test serum 
aliquots (for a final dilution of 1:4) 
5. For a full run (i.e. 96 sera), label two stacks of 12 microplates for each serotype (Figure 3). 
a. Use a lidded microplate for the top of each stack, with the low evaporation lid  
b. The 11 remaining microplates can be lidded or lidless; removal of lids is required for many 
automation systems 
c. Cell control and back titration plates can be set up in lidded plates 
 
 
 
Plate number 
Run number 
P1 plate number 

 
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AUTOMATED 
A6.  Use the MicroFlo reagent dispenser program “MEDIA 25UL” to add 25 μl MEM 2% FBS to the test 
plates.  
a. This program adds 25 μl MEM+2% FBS to all wells for each test plate and the back titration 
plates (PV1, PV2, and PV3).  
b. For the cell control plate, a total of 50 μl MEM+2% FBS should be added to each well. This 
plate can be filled twice using the “MEDIA 25UL” program. 
A7.  Use the TECAN programs “1 or 2 plate triplicate transfer” to make triplicate sample plates (Figures 
4A). 
a. This program will transfer 100 µL of test serum in a deep-well plate (from Step 3) in 
triplicate to a triplicate sample plate. 
A8.  Using electronic single-channel pipette, transfer 25 μL of test serum from triplicate sample plate (Step 
6) to three different test plates (one for each serotype) (Figure 4B). 
 
A 
 
96-Deep Well Plate 
 
 
 
 
Triplicate Sample Plate 
 
 
B 
 
Triplicate Sample Plate  
 
 
 
 
Test Plate 
 
 
Figure 4. . For each run, 100 µl of sample is transferred from sample plate to dilution plates in triplicate (enough 
for starting three 25 µl sample dilutions) (A). For simplicity, only the first 8 samples are shown in the deep 96--
well plate. The TECAN configuration will allow for triplicates in consecutive order. This step can transfer 96 
samples from one 96-deep well plate to a 96-well triplicate sample plate in 22 min. (B) Using a 12-channel manual 
pipette, each row of from the triplicate sample plate is transferred to a test plate in triplicate for each serotype. 
 
A9.  Use the WellPro program “SD25” to perform 2-fold serial dilutions (Figure 5). 
a. This program will transfer 25 μL of the diluted serum in row A to row B, then from row B to 
row C, etc. 

 
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b. For row H, 25 μL will be aspirated and discarded with the tips, to maintain the correct total 
volume. 
c. For each transfer, the program will mix the diluted serum a total of 4 times. 
d. Using this program will produce serum dilutions ranging from 1:8 to 1:1024. 
e. This step generates the test plates, ready for addition of virus and cells. 
 
 
 
               Test Plate 
 
 
 
    
 
     Diluted Test Plate 
 
Figure 5. Each test serum is serially diluted (2-fold) from 1:8 to 1:1024 in triplicate. This is repeated for each test 
plate designated for poliovirus serotypes 1, 2, or 3.  
 
A10.  Arrange the plates in stacks of 12-13; use a lid to cover the top plate of each stack of plates and wrap 
in plastic Saran wrap. 
A11.  Plates can be stored overnight at 4°C. 
 
MANUAL 
M5.  Use electronic multichannel pipette to dispense 25 μL MEM+2% FBS to the test plates.  
a. Add 25 μl MEM+2% FBS to each well of the back titration plates (PV1, PV2, and PV3). 
b. Add 50 μl MEM+2% FBS to each well of the cell control plate. 
M6.  Use an electronic multichannel pipette to transfer 25 μL of each test serum (from Step 3) to each test 
plate in triplicate (Figure 4B). 
M7.  Using a multichannel pipette, make serial 2-fold dilutions from row A to row H. (serum dilution will 
range from 1:8 to 1:1024) (see Figure 5 for plate layout). 
a. Discard 25 μL from row H to make final volume for all wells 25 μL. 
M8.  Cover the top plate of each stack of plates and wrap in plastic wrap. 
M9.  Plates can be stored overnight at 4°C. 
 
 Day of run: 
 
1. Dilute each virus in MEM+2% FBS to contain 100 CCID50 according to Table 2.  
a. Prepare sufficient virus challenge suspension for the number of sera to be tested; each plate 
requires approximately 2.5 ml of diluted challenge virus. 
2. Prepare the back titrations of each poliovirus serotype in MEM+2% FBS. Titrate each virus from 100 
CCID50 a further three 10-fold steps (Figure 6). 
 
 
 

 
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0.1 TCID50 
100 μL 
900 μL 
 
100 μL 
900 μL 
 
100 μL 
900 μL 
Back Titration Plate 
1 TCID50 
100 μL 
900 μL 
 
100 μL 
900 μL 
 
100 μL 
900 μL 
10 
TCID50 
100 μL 
900 μL 
 
100 μL 
900 μL 
 
100 μL 
900 μL 
100 
TCID50 
7 mL 
63 mL 
 
7 mL 
63 mL 
 
7 mL 
63 mL 
Working 
Stock 
 
800 μL 
7.2 mL 
 
800 μL 
7.2 mL 
 
800 μL 
7.2 mL 
 
 
100 μL 
900 μL 
 
100 μL 
900 μL 
 
 
 
 
 
100 μL 
900 μL 
 
100 μL 
900 μL 
 
100 μL 
900 μL 
 
Sabin 1 (NIBSC 01/528) 
100 TCID50 = 10-5.28 
100 μL 
900 μL 
Sabin 2 (NIBSC 01/530) 
100 TCID50 = 10-5.2 
100 μL 
900 μL 
Sabin 3 (NIBSC 01/532) 
100 TCID50 = 10-4.68 
100 μL 
900 μL 
 
100 μL 
38 μL 
100 μL 
82 μL 
100 μL 
82 μL 
 
Virus 
Medium 
Virus 
Medium 
Virus 
Medium 
 
 
 
 
 
Table 2. Example Virus Dilution for 100 TCID50 and Back Titration Plate for Sabin type 1, 2, and 3 

 
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Figure 6. Layout for back titration plate. A back titration plate is made for each virus tested in the 
microneutralization assay. 
 
AUTOMATED 
A3.    Use the MicroFlo program VIRUS 25UL to add 25 μl of 100 CCID50 of relevant poliovirus antigen 
to all wells in the diluted serum test plates. Use a different sterile dispensing cartridge for each 
virus. 
 
MANUAL 
M3.  Use an electronic, multichannel pipette to add 25 µL of 100 CCID50 of relevant poliovirus antigen 
to all wells in the diluted test plates. 
M4.  Prepare back titration plate (Figure 6) for each Sabin strain using dilutions prepared in step 2 (Table 
2). 
a. Add 25 μl of 100 TCID50 of virus to rows A and B (i.e., 24 wells/dilution) 
b. Add 25 μl of the next three 10-fold dilutions to rows C and D, E and F, and G and H, 
respectively (See Table 2 for dilutions). 
c. Change pipette tips between each dilution 
M5.  Wrap all plates in plastic wrap and incubate for 3 hours at 35°C and 5% CO2 
M6.  During serum-virus incubation, wash HEp2-C monolayer cell cultures (from 150 cm2 flasks), 
trypsinize, and count cells using automatic cell counter (BioRad) or a hemocytometer. 
M7.  Prepare a cell suspension in MEM+10% FBS to contain 3 x 105 cells/ml. Prepare a sufficient volume 
of cells: each plate requires approximately 2.5 ml of cell suspension, and every run requires 3-4 
confluent 150 cm2 flasks. Store cells in glass bottle at 4°C until ready to use. 
 
AUTOMATED 
A4.   Use MicroFlo program “CELL 25UL” to add 25 μl of prepared cell suspension to each well of every 
plate.  
 
MANUAL 

 
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M8.  Use a repeating, multichannel pipette to add 25 μL of prepared cell suspension to each well of 
every plate. 
M9.  Wrap all plates in plastic wrap, in stacks of 12-13 plates. To prevent spills and cross-contamination, 
avoid abrupt handling of plates. 
M10. Carefully transfer plates to incubator for 5 days incubation at 35°C and 5% CO2.  
 
Plate washing and staining. 
 
AUTOMATED 
A1.   After 5 days incubation, use the Biotek EL406 to wash and stain plates with crystal violet solution. 
A2.  This will aspirate wells and dispense 50 μL of crystal violet stain (0.05%) to all wells. 
A3.  Incubate for a minimum of 40 minutes at room temperature. 
A4.  Run wash program 
a. This program will aspirate the crystal violet stain then fill all wells with 250 μL of tap water and 
aspirate. 
b. This process will be repeated 3 more times to completely remove any excess crystal violet stain 
in the test plates prior to reading. 
 
MANUAL 
M1.  After 5 days incubation, aspirate/discard media with multichannel pipette into freshly made 0.5% 
sodium hypochlorite solution.  
M2.  Using a repeating, multichannel pipette, add 50 μL crystal violet stain (0.05%) to all plates. 
M3.  Incubate for a minimum of 40 minutes at room temperature. 
M4.  Aspirate/discard stain with multichannel pipette, fill each well with tap water (approximately 250-
300 μL), and discard. 
a. Repeat washing step 3 more times. 
M5.  Allow plates to dry at room temperature for approximately 30 minutes.  
M6.  Plates should be kept at room temperature until results are calculated and results are reported. 
 
Notes 
 
1. Data Collection 
 
AUTOMATED 
A1.  Read plates with ELISA reader at 595 nm wavelength. 
a. Data from plate reader can be processed using macros written in Visual Basic for Microsoft 
Excel. 
b. Use an absorbance cutoff value of ≥ 0.8 as positive for neutralization (i.e., intact monolayer). 
c. Automatically calculate the titer for each specimen using Formula 1 below (Step 2). 
 
MANUAL 
M1.  For each triplicate test serum, count the total number of wells positive for neutralization    (i.e., 
purple wells). 
M2.  To calculate titer: 
 
Formula 1 
Titer = (# positive wells / # replicates) + 2.5 
 

 
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a. To calculate reciprocal titer: 
 
Formula 2 
Reciprocal titer = 2titer 
 
M3.  For the neutralization titers, the upper limit of detection is 10.5 and the lower limit is 2.5, which is 
considered negative. 
 
2. Cross-checking stained plates 
 
1. To ensure accuracy, each plate is cross checked to verify correct order of plates and compare plate 
staining pattern to electronic data file to verify titer.  
2. Due to the variance of the assay, there is a statistical likelihood that some wells will have virus not 
neutralizied by antibody despite neutralization at lower serum dilutions. 
3. To account for this, stained plates should be checked for these situations and the data corrected 
according to the following guidelines (Figure 7). 
 
 
 
Figure 7. Example of cross-checking a crystal violet stained 96-well plate for poliovirus serology assay. 
 
Example 1 
a. For Test Serum 1, the reader software will automatically read this as a titer of 7.83 (1:227) 
b. By cross-checking the plate, we will count D1 as positive for neutralization because the 1:128 
and 1:256 dilutions (E1 and F1) are positive for neutralization (i.e., purple). 
c. Using Formula 1, the titer is adjusted to 8.17 (1:288) 
 
Example 2 
a. For Test Serum 2, the reader software will automatically read this as a titer of 6.83 (1:114) 
b. By cross-checking the plate, we will count F5 as negative for neutralization because the 1:64 
dilutions (D4, D5, and D6) are negative for viral growth 

 
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3.  Summary of Randomization Process 
 
1. For each study, the specimen number, study site, study arm, patient number, and the unique ID for each 
specimen. 
2. Each run needs to have 4-6 in-house reference sera.  
a. Based on the total number of runs and the number of divisions per plate, the program will also 
determine how many plates will be required. 
b. The function determines how many unique subjects exist in the file and how many unique study 
groups exist (where a study group is a unique combination of study site and study arm for multi-
site or multi-arm studies). Each study group will be evenly represented in each run, so the 
program assigns the subjects within each study group randomly and evenly amongst the runs. 
c. After each subject identifier is assigned a run number, all sera associated with those subject 
identifiers are assigned to the same run. 
d. Finally, plate numbers and plate positions are randomly assigned to the sera within each run, and 
a spreadsheet of the randomized sera is created, in MS Excel format. 
3. At the prompt, enter in the first run number for this group of serology runs. 
4. At the next prompt, enter in the code that represents the first specimen per subject (for example SE1) in 
the input file. 
5. At the next prompt, select the path and name of the output file. 
6. A dialog box will appear when the process is complete. 
7. Review the output to confirm that all samples were included in the randomized output. 
 
4. In-House Reference Serum (IHRS) 
 
1. The in-house reference serum should be established by measuring polio neutralizing antibody titers in a 
population of immunized subjects.  
2. Multiple sera with high neutralization titers (≥ 7.5) should be pooled to generate a high-volume control. 
3. Make 100 μL aliquots of the pooled sera to be stored at -20°C for future use. Once thawed, IHRS 
aliquots must be stored at 4°C for no more than 3-4 days. 
4. To prepare the IHRS for use in the serology assay, adjust the initial dilution such that the endpoint titer 
is reached on the 3-5 dilution on an assay plate.  
Make the initial dilution of the IHRS and transfer to the wells that are designated for the IHRS in the 
randomized list (See Table 1).  
 
 

 
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Appendix C: Summary of Laboratory Methods for Stool Testing 
 
Stool samples will be stored at -20°C until analysis. Poliovirus genomes will be detected using a 
Sabin multiplex real-time RT-PCR (rRT-PCR) assay of total nucleic acid extracted from stool 
suspensions (50%, w/v) [14]. Virus titrations will be performed as described previously [14]. For stool 
samples with >105 CCID50 per ml, stool suspensions (10% w/v in EMEM) will be prepared for high 
resolution sequencing. For stools with low Ct values in Sabin 2 realtime PCR assays, a subset of the 
stool samples will be subjected to virus isolation/expansion prior to RNA extraction and high resolution 
sequencing. Virus will be amplified in HEp-2C (ATCC, CCL-23) cells maintained in EMEM 
supplemented with 2% fetal bovine serum (Atlanta Biologicals). Confluent monolayers of HEp-2C cells 
in 24-well cell culture cluster plates (Costar) will be inoculated with 50 µL of each 10% stool 
suspension (4 wells, at least 50 CCID50 per well) and incubated for 3 days at 33°C, 5% CO2. Following 
two freeze-thaw cycles, cell lysate will be pooled from the 4 wells, and virus isolates will be harvested 
following removal of cell debris by centrifugation at 3000 x g for 10 minutes.  
Sequence and data analysis will be performed on viral RNA isolated from both culture-amplified 
virus and from 10% stool suspensions, using a previously described methods [14]. In brief, viral RNA 
will be isolated from 140 µl amplified virus stock or stool suspension using a QIAamp Viral RNA mini 
kit (Qiagen). cDNA preparation and first round amplification of nearly-full-length genome will be 
performed using poliovirus-specific primers near the ends of the genome, followed by semi-nested PCR 
using a poliovirus-specific primer at the 5’ end of the genome and a Sabin 2-specific primer in the 
carboxy terminus of VP1. The other half of the genome will be amplified by Sabin 2-specific primer at 
the amino terminus of VP1 and poliovirus-specific primer at the 3’ end of the genome.  Library 
preparation by Nextera XT (Illumina) and paired-end sequencing using MiSeq reagents on a MiSeq 
instrument to generate FASTQ files.  
Data analysis will be performed using the appropriate reference sequence and sequence analysis 
pipeline. Sequencing reads will be mapped against all three Sabin vaccine reference sequences to 

 
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generate coverage plots. As needed, amplicons will be analyzed using long-read sequencing methods 
(MinIon or PacBio), to confirm the genome structure.  
 
 
 

 
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Appendix D: External Reviews 
 
Reviewer 1 
 
Name of Reviewer: Ondrej Mach   
Position: POL Research team Lead 
Institution: WHO 
 
EVALUATION FORM 
 
Title:  
 
Summary of Referee's Opinions: 
 
 
 
 
Rank Score 
 
 
High 
Medium  Low 
Quality of project 
X 
 
 
Adequacy of project design  
X 
 
 
Suitability of methodology 
X 
 
 
Feasibility within time period 
X 
 
 
Appropriateness of budget 
 
X 
 
Potential value of field of knowledge 
X 
 
 
  
 
CONCLUSIONS                                                       
 
I support the project proposal 
 
a) without qualification 
X 
b)  with qualification 
 
c)  on technical grounds 
 
d)  on level of financial support 
 
                                                  
I do not support the project proposal 
 
 
Name of Reviewer: _________Ondrej Mach__________________   
 
Signature:__________________ 
 
 
.Date:___20/03/2020____________ 
Position: ___POL Research team Lead______________________________ 
Institution:  ___WHO____________________________________ 
 
Detailed Comments: (Please use additional page if necessary.) 
 
This is an important study that will provided much needed additional clinical trial 
data for nOPV and first data on co-administration of  nOPV and bOPV. I fully 
support this study. 
 
Response: Thanks 
 
 

 
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Reviewer 2 
 
Name: Professor Dr. Md. Shafiqul Islam 
Position: Former Professor and Head, Department of Epidemiology, 
Institution: National Institute of Preventive and Social Medicine (NIPSOM). 
 
Title:  Immunogenicity of novel monovalent oral poliovirus vaccine type 2 (nOPV2) with and without 
bivalent OPV 
 
Summary of Referee’s Opinions: Please see the following table to evaluate the various aspects of the 
proposal by checking the appropriate boxes. Your detailed comments are sought on a separate, attached 
page. 
 
 
Rank Score 
 
High 
Medium 
Low 
Quality of project 
√ 
 
 
Adequacy of project design 
√ 
 
 
Suitability of methodology 
√ 
 
 
Feasibility within time period 
√ 
 
 
Appropriateness of budget 
√ 
 
 
Potential value of field of knowledge 
√ 
 
 
 
CONCLUSIONS 
 
I support the application: 
 
a)  without qualification 
 
√ 
 
b)  with qualification  
 
 
      - on technical grounds 
 
 
 
      -  on level of financial support 
 
 
I do not support the application 
 
 
 
 
Name of Referee:  Professor Dr. Md. Shafiqul Islam 
 
 
Signature :    
 
 
 
 
 
Date : 13/04/2020 
Position: Former Professor and Head, Department of Epidemiology, 
 
Institution: National Institute of Preventive and Social Medicine (NIPSOM). 
 

 
119 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Detailed Comments 
Please briefly provide your opinions of this proposal, giving special attention to the originality and 
feasibility of the project, its potential for providing new knowledge and the justification of financial 
support sought; include suggestions for modifications (scientific or financial) where you feel they are 
justified. (Use additional pages if necessary) 
 
Title: Immunogenicity of novel monovalent oral poliovirus vaccine type 2 (nOPV2) with and without 
bivalent OPV 
PI : Dr. K Zaman 
Name of Reviewer : Prof. Dr. Md. Shafiqul Islam 
 
Comments:   
Wild poliovirus type 2 (WPV2) was declared eradicated in 2015 and type 2 oral poliovirus vaccine (OPV2) was 
withdrawn globally in May 2016. Monovalent OPV2 (mOPV2) is stockpiled for responding to circulating type 2 
vaccine-derived poliovirus (cVDPV2) outbreaks. However, Sabin polioviruses in OPV can revert to 
neurovirulence and there is evidence that mOPV2 vaccines used to control cVDPV2 outbreaks have generated 
new VDPV2 emergences that have led to cVDPV2 outbreaks. Novel OPV2 (nOPV2) candidates have been 
developed with improved genetic stability to reduce the risk of seeding new VDPVs. Findings from a phase 1 
clinical trial in adults demonstrated the safety and immunogenicity of two nOPV2 candidates. Preliminary results 
from phase 2 clinical trials in adults and children also suggest that the nOPV2 vaccines are safe and 
immunogenic. It is expected that nOPV2 would replace mOPV2 for responding to type 2 outbreaks. Outbreak 
response for cVDPV2 also offers the opportunity to close immunity gaps to polioviruses types 1 and 3. It is 
important to get data on the immunogenicity of co-administered nOPV2 and bOPV, compared to either vaccine 
given alone. No study has yet assessed the immunogenicity of co-administered nOPV2 and bOPV. So, this is an 
important study to fill up this knowledge gap.  
  
This is an open-label, non-inferiority, inequality balanced five arm randomized clinical trial assessing 
immunogenicity of poliovirus vaccines. Vaccine recipient will be enrolled and randomized at 6 weeks to one of 
five arms:  
A: nOPV2 candidate 1 @ 6, 10, and 14 weeks  
B: nOPV2 candidate 1 + bOPV @ 6, 10, and 14 weeks  
C: nOPV2 candidate 2 @ 6, 10, and 14 weeks  
D: nOPV2 candidate 2 + bOPV @ 6, 10, and 14 weeks  
E: bOPV @ 6, 10, and 14 weeks  
 
This is a well written research protocol. The investigators as well as this prestigious organization have long 
experience of conducting vaccine trial research project. This research project has the potential to play an 
important role to guide Global Polio Eradication Initiative (GPEI), including WHO,  in making policy for 
responding to polio outbreaks with the goal of eradication of polio worldwide. 
 
 
Prof. Dr. Md. Shafiqul Islam 
Former Professor and Head, Department of Epidemiology, 
NIPSOM, 
Mohakhali, Dhaka 1212. 
 
 
Response: Thanks  
 

 
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Check-List 
 
Check-list for Submission of Research Protocol 
For Consideration of the Research Review Committee (RRC) 
[Please check all appropriate boxes] 
 
1. Has the proposal been reviewed, discussed and cleared by all listed investigators? 
 
  Yes                
  No 
If the response is No, please clarify the reasons:       
 
2. Has the proposal been peer-reviewed externally?  
 
  Yes               
  No                    
  External Review Exempted 
If the response is ‘No’ or “External Review Exempted”, please explain the reasons:       
 
 
If the response is “Yes”, please indicate if all of their comments have been addressed? 
  Yes (please attach)              
 
  No (please indicate reason(s)): 
 
3. Has the budget been reviewed and approved by icddr,b’s Finance? 
  Yes                
 No (reason): ________________________________________________________________ 
4. Has the Ethics Certificate(s) been attached with the Protocol? 
 
  Yes                
  No  
If the answer is ‘No’, please explain the reasons:       
 
 
 
 
 
_______________________________                                                                                                        _________            
Signature of the Principal Investigator 
                                                                                                    Date             
                                                               
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
121 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
E-mail approvals of Investigators 
 
From: Dr. Mohammad Yunus  
Sent: Wednesday, June 24, 2020 8:37 AM 
To: Dr. K. Zaman <kzaman@icddrb.org>; Asma Binte Aziz <asma.aziz@icddrb.org>; Masuma Hoque 
<masuma.hoque@icddrb.org> 
Subject: RE: Approval for submission of nOPV+BOPV protocol 
 
Dear Dr. Zaman, 
I approve the protocol entitled “Immunogenicity of novel monovalent oral poliovirus vaccine type 2 
(nOPV2) with and without bivalent OPV" for submission to RRC. 
Thank you very much and best wishes. 
Md. Yunus 
 
 
 
From: Dr. K. Zaman  
Sent: Tuesday, June 23, 2020 8:31 PM 
To: Dr. Mohammad Yunus; Asma Binte Aziz; Masuma Hoque 
Subject: Approval for submission of nOPV+BOPV protocol 
 
Dear All, 
We need your approval for submission to  the RRC of the following protocol : 
“Immunogenicity of novel monovalent oral poliovirus vaccine type 2 (nOPV2) with and without 
bivalent OPV". 
 
Please find attached the protocol. 
Appreciate if you please give it urgently (planning to submit tomorrow). 
 
Best regards 
Zaman 
 
 
From: Asma Binte Aziz  
Sent: Tuesday, June 23, 2020 8:34 PM 
To: Dr. K. Zaman <kzaman@icddrb.org> 
Cc: Dr. Mohammad Yunus <myunus@icddrb.org>; Masuma Hoque <masuma.hoque@icddrb.org> 
Subject: Re: Approval for submission of nOPV+BOPV protocol 
 
Dear Sir, 
Thanks. 
Please proceed for RRC submission. 
 
Best Regards, 
Asma 
 
On 23 Jun, 2020, at 8:31 PM, Dr. K. Zaman <kzaman@icddrb.org> wrote: 
 
Dear All, 
We need your approval for submission to  the RRC of the following protocol : 

 
122 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
“Immunogenicity of novel monovalent oral poliovirus vaccine type 2 (nOPV2) with and without 
bivalent OPV". 
  
Please find attached the protocol. 
Appreciate if you please give it urgently (planning to submit tomorrow). 
  
Best regards 
Zaman 
  
 
From: Masuma Hoque  
Sent: Tuesday, June 23, 2020 8:44 PM 
To: Dr. K. Zaman 
Cc: M.A Salam Khan 
Subject: Approval  
 
Zaman Bhai,  
 
I approve  submission of the following protocol  to  the RRC : 
 
“Immunogenicity of novel monovalent oral poliovirus vaccine type 2 (nOPV2) with and without 
bivalent OPV". 
  
Sincerely 
Masuma 
 
 
From: Wilkinson, Amanda (CDC/DDPHSIS/CGH/GID) [mailto:lxq6@cdc.gov]  
Sent: Wednesday, June 10, 2020 6:58 PM 
To: Dr. K. Zaman <kzaman@icddrb.org> 
Cc: Masuma Hoque <masuma.hoque@icddrb.org>; Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) 
<dvi5@cdc.gov>; Snider, Cynthia J. (CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Asma Binte Aziz 
<asma.aziz@icddrb.org> 
Subject: nOPV2 + bOPV trial protocol  
 
Dear Dr. Zaman, 
 
Attached is an updated nOPV2 + bOPV trial protocol. 
 
The following items need to be added, but otherwise I think this is ready for RRC submission.  
1. Translated consent form 
2. Second external reviewer form 
 
Please let me know if there’s anything else you need from our side.  
Best wishes, 
Amanda 
 
From: Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) [mailto:dvi5@cdc.gov]  
Sent: Monday, June 22, 2020 8:12 AM 
To: Dr. K. Zaman <kzaman@icddrb.org>; Snider, Cynthia J. (CDC/DDPHSIS/CGH/GID) 
<bex4@cdc.gov>; Estivariz, Concepcion (Conchi) (CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, 

 
123 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Qian (CDC/DDPHSIS/CGH/GID) <fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) 
<map1@cdc.gov>; Wassilak, Steve (CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Oberste, Steve 
(CDC/DDID/NCIRD/DVD) <mbo2@cdc.gov>; Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) 
<wxx3@cdc.gov> 
Subject: RE: nOPV2 + bOPV trial protocol  
 
Dear Zaman, 
I approve the submission to RRC. 
 
Best regards, 
Abhijeet 
 
From: Dr. K. Zaman <kzaman@icddrb.org>  
Sent: Saturday, June 20, 2020 2:04 PM 
To: Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov>; Snider, Cynthia J. 
(CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, Concepcion (Conchi) 
(CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, Qian (CDC/DDPHSIS/CGH/GID) 
<fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) <map1@cdc.gov>; Wassilak, Steve 
(CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Oberste, Steve (CDC/DDID/NCIRD/DVD) 
<mbo2@cdc.gov>; Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: nOPV2 + bOPV trial protocol  
 
Dear All, 
My sincere thanks to you all for reviewing the protocol “Immunogenicity of novel monovalent oral 
poliovirus vaccine type 2 (nOPV2) with and without bivalent OPV”. We need your approval for 
submission to our Research Review Committee. 
Requesting to write “Approve for submission”. 
 
Kindest regards 
Zaman 
 
Dr. K. Zaman 
Senior Scientist, Enteric and Respiratory Infections 
Infectious Diseases  Division 
 
 
From: Snider, Cynthia J. (CDC/DDPHSIS/CGH/GID) [mailto:bex4@cdc.gov]  
Sent: Monday, June 22, 2020 6:11 PM 
To: Dr. K. Zaman <kzaman@icddrb.org>; Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) 
<dvi5@cdc.gov>; Estivariz, Concepcion (Conchi) (CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, 
Qian (CDC/DDPHSIS/CGH/GID) <fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) 
<map1@cdc.gov>; Wassilak, Steve (CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Oberste, Steve 
(CDC/DDID/NCIRD/DVD) <mbo2@cdc.gov>; Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) 
<wxx3@cdc.gov> 
Subject: RE: nOPV2 + bOPV trial protocol  
 
Dear Zaman, 
I approve for submission. 
Best regards, 

 
124 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Cindi 
 
 
From: Dr. K. Zaman <kzaman@icddrb.org>  
Sent: Saturday, June 20, 2020 2:04 PM 
To: Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov>; Snider, Cynthia J. 
(CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, Concepcion (Conchi) 
(CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, Qian (CDC/DDPHSIS/CGH/GID) 
<fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) <map1@cdc.gov>; Wassilak, Steve 
(CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Oberste, Steve (CDC/DDID/NCIRD/DVD) 
<mbo2@cdc.gov>; Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: nOPV2 + bOPV trial protocol  
 
Dear All, 
My sincere thanks to you all for reviewing the protocol “Immunogenicity of novel monovalent oral 
poliovirus vaccine type 2 (nOPV2) with and without bivalent OPV”. We need your approval for 
submission to our Research Review Committee. 
Requesting to write “Approve for submission”. 
 
Kindest regards 
Zaman 
 
Dr. K. Zaman 
Senior Scientist, Enteric and Respiratory Infections 
Infectious Diseases  Division 
 
 
From: Estivariz, Concepcion (Conchi) (CDC/DDPHSIS/CGH/GID) [mailto:cge3@cdc.gov]  
Sent: Monday, June 22, 2020 7:01 PM 
To: Dr. K. Zaman <kzaman@icddrb.org> 
Subject: RE: nOPV2 + bOPV trial protocol  
 
Dear Zaman 
I approve for submission.  
 
Best Regards 
Conchi 
From: Dr. K. Zaman <kzaman@icddrb.org>  
Sent: Saturday, June 20, 2020 2:04 PM 
To: Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov>; Snider, Cynthia J. 
(CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, Concepcion (Conchi) 
(CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, Qian (CDC/DDPHSIS/CGH/GID) 
<fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) <map1@cdc.gov>; Wassilak, Steve 
(CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Oberste, Steve (CDC/DDID/NCIRD/DVD) 
<mbo2@cdc.gov>; Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: nOPV2 + bOPV trial protocol  
 
Dear All, 

 
125 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
My sincere thanks to you all for reviewing the protocol “Immunogenicity of novel monovalent oral 
poliovirus vaccine type 2 (nOPV2) with and without bivalent OPV”. We need your approval for 
submission to our Research Review Committee. 
Requesting to write “Approve for submission”. 
 
Kindest regards 
Zaman 
 
Dr. K. Zaman 
Senior Scientist, Enteric and Respiratory Infections 
Infectious Diseases  Division 
 
From: An, Qian (CDC/DDPHSIS/CGH/GID) [mailto:fei8@cdc.gov]  
Sent: Wednesday, June 24, 2020 12:39 AM 
To: Wassilak, Steve (CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Dr. K. Zaman 
<kzaman@icddrb.org>; Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov>; Snider, 
Cynthia J. (CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, Concepcion (Conchi) 
(CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) 
<map1@cdc.gov>; Oberste, Steve (CDC/DDID/NCIRD/DVD) <mbo2@cdc.gov>; Lickness, Jacquelyn 
S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: RE: nOPV2 + bOPV trial protocol  
 
Dear Dr. Zaman, 
 
I approve for the submission. 
Thanks, 
Qian 
 
From: Pallansch, Mark A. (CDC/DDID/NCIRD/OD) [mailto:map1@cdc.gov]  
Sent: Sunday, June 21, 2020 10:54 PM 
To: Dr. K. Zaman <kzaman@icddrb.org>; Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) 
<dvi5@cdc.gov>; Snider, Cynthia J. (CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, 
Concepcion (Conchi) (CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, Qian 
(CDC/DDPHSIS/CGH/GID) <fei8@cdc.gov>; Wassilak, Steve (CDC/DDPHSIS/CGH/GID) 
<sgw1@cdc.gov>; Oberste, Steve (CDC/DDID/NCIRD/DVD) <mbo2@cdc.gov>; Lickness, Jacquelyn 
S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: RE: nOPV2 + bOPV trial protocol  
 
I approve for submission. 
 
MAP 
 
 
From: Dr. K. Zaman <kzaman@icddrb.org>  
Sent: Saturday, June 20, 2020 2:04 PM 
To: Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov>; Snider, Cynthia J. 
(CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, Concepcion (Conchi) 
(CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, Qian (CDC/DDPHSIS/CGH/GID) 
<fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) <map1@cdc.gov>; Wassilak, Steve 
(CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Oberste, Steve (CDC/DDID/NCIRD/DVD) 

 
126 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
<mbo2@cdc.gov>; Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: nOPV2 + bOPV trial protocol  
 
Dear All, 
My sincere thanks to you all for reviewing the protocol “Immunogenicity of novel monovalent oral 
poliovirus vaccine type 2 (nOPV2) with and without bivalent OPV”. We need your approval for 
submission to our Research Review Committee. 
Requesting to write “Approve for submission”. 
 
Kindest regards 
Zaman 
 
Dr. K. Zaman 
Senior Scientist, Enteric and Respiratory Infections 
Infectious Diseases  Division 
 
 
 
From: Wassilak, Steve (CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>  
Sent: Tuesday, June 23, 2020 2:07 PM 
To: Dr. K. Zaman <kzaman@icddrb.org>; Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) 
<dvi5@cdc.gov>; Snider, Cynthia J. (CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, 
Concepcion (Conchi) (CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, Qian 
(CDC/DDPHSIS/CGH/GID) <fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) 
<map1@cdc.gov>; Oberste, Steve (CDC/DDID/NCIRD/DVD) <mbo2@cdc.gov>; Lickness, Jacquelyn 
S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: RE: nOPV2 + bOPV trial protocol  
 
Dear Dr Zaman 
I approve for  submission 
 
Best 
SW 
 
Steven Wassilak MD 
Global Immunization Division, CDC, H24-2 
Tel: +1 404 639 1867  
Mob: +1 770 331 7695 
email: swassilak@cdc.gov or sgw1@cdc.gov 
 
 
From: Oberste, Steve (CDC/DDID/NCIRD/DVD) [mailto:mbo2@cdc.gov]  
Sent: Sunday, June 21, 2020 7:18 AM 
To: Dr. K. Zaman <kzaman@icddrb.org>; Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) 
<dvi5@cdc.gov>; Snider, Cynthia J. (CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, 
Concepcion (Conchi) (CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, Qian 
(CDC/DDPHSIS/CGH/GID) <fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) 
<map1@cdc.gov>; Wassilak, Steve (CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Lickness, 
Jacquelyn S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: RE: nOPV2 + bOPV trial protocol  

 
127 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
 
Zaman: 
 
I approve for submission. 
 
Steve 
 
From: Dr. K. Zaman <kzaman@icddrb.org>  
Sent: Saturday, June 20, 2020 2:04 PM 
To: Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov>; Snider, Cynthia J. 
(CDC/DDPHSIS/CGH/GID) <bex4@cdc.gov>; Estivariz, Concepcion (Conchi) 
(CDC/DDPHSIS/CGH/GID) <cge3@cdc.gov>; An, Qian (CDC/DDPHSIS/CGH/GID) 
<fei8@cdc.gov>; Pallansch, Mark A. (CDC/DDID/NCIRD/OD) <map1@cdc.gov>; Wassilak, Steve 
(CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Oberste, Steve (CDC/DDID/NCIRD/DVD) 
<mbo2@cdc.gov>; Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) <wxx3@cdc.gov> 
Subject: nOPV2 + bOPV trial protocol  
 
Dear All, 
My sincere thanks to you all for reviewing the protocol “Immunogenicity of novel monovalent oral 
poliovirus vaccine type 2 (nOPV2) with and without bivalent OPV”. We need your approval for 
submission to our Research Review Committee. 
Requesting to write “Approve for submission”. 
 
Kindest regards 
Zaman 
 
Dr. K. Zaman 
Senior Scientist, Enteric and Respiratory Infections 
Infectious Diseases  Division 
 
 
From: Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) [mailto:wxx3@cdc.gov]  
Sent: Tuesday, June 23, 2020 10:36 PM 
To: Dr. K. Zaman <kzaman@icddrb.org>; An, Qian (CDC/DDPHSIS/CGH/GID) <fei8@cdc.gov>; 
Wassilak, Steve (CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov> 
Cc: Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov> 
Subject: RE: nOPV2 + bOPV trial protocol  
 
Dear Dr. Zaman, 
I approve for submission. 
Thank you, 
 
J. Sunshine Lickness, MPH | Surveillance, Innovation and Research Team – Global Immunization Division – Center for Global Health – 
US Centers for Disease Control and Prevention | Tel (office): 404-639-8039 | Mobile: 678-519-8389 | E-mail: wxx3@cdc.gov  
 
From: Dr. K. Zaman <kzaman@icddrb.org>  
Sent: Tuesday, June 23, 2020 12:32 PM 
To: An, Qian (CDC/DDPHSIS/CGH/GID) <fei8@cdc.gov>; Wassilak, Steve 
(CDC/DDPHSIS/CGH/GID) <sgw1@cdc.gov>; Lickness, Jacquelyn S. (CDC/DDPHSIS/CGH/GID) 
<wxx3@cdc.gov> 

 
128 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
Cc: Anand, Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov> 
Subject: RE: nOPV2 + bOPV trial protocol  
Importance: High 
 
Dear All, 
I am not sure if I missed your mail.  
My sincere thanks to you all for reviewing the protocol “Immunogenicity of novel monovalent oral 
poliovirus vaccine type 2 (nOPV2) with and without bivalent OPV”.  
We need your approval for submission to our Research Review Committee. 
Requesting to write “Approve for submission”. 
 
Kindest regards 
Zaman 
 
Dr. K. Zaman 
Senior Scientist, Enteric and Respiratory Infections 
Infectious Diseases  Division 
 
 
----------------------------------------- 
From: Wilkinson, Amanda (CDC/DDPHSIS/CGH/GID) <lxq6@cdc.gov>  
Sent: Monday, November 23, 2020 9:14 AM 
To: Patel, Jaymin (CDC/DDPHSIS/CGH/GID) <isr0@cdc.gov>; Anstadt, Jennifer 
(CDC/DDID/NCIRD/DVD) <yrq1@cdc.gov>; Burns, Cara C. (CDC/DDID/NCIRD/DVD) 
<zqd1@cdc.gov>; Ananda Bandyopadhyay <Ananda.Bandyopadhyay@gatesfoundation.org>; Jillian 
Gauld <jgauld@idmod.org>; Mike Famulare <mfamulare@idmod.org> 
Cc: Dr. K. Zaman <kzaman@icddrb.org>; Masuma Hoque <masuma.hoque@icddrb.org>; Anand, 
Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov> 
Subject: Approval of nOPV2-bOPV protocol from  
 
Dear all, 
 
Thanks very much for your contributions to the protocol “Immunogenicity of novel monovalent oral 
poliovirus vaccine type 2 (nOPV2) with and without bivalent OPV”. Revised versions of the protocol 
are attached.  
 
Icddr,b needs your approval for submission to their Research Review Committee. If you approve, please 
respond to this email with “Approve for submission”. 
 
Best wishes, 
Amanda 
 
From: Patel, Jaymin (CDC/DDPHSIS/CGH/GID) <isr0@cdc.gov>  
Sent: Monday, November 23, 2020 9:17 AM 
To: Wilkinson, Amanda (CDC/DDPHSIS/CGH/GID) <lxq6@cdc.gov> 
Subject: RE: Approval of nOPV2-bOPV protocol from 
 
Approve for submission 
 

 
129 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
__________________________________ 
Jaymin Patel, PhD, MPH  
LCDR, U.S. Public Health Service 
Epidemiologist, Global Immunization Division 
Centers for Disease Control and Prevention 
Office: (404) 718-5539 | Cell: (252) 347-1908 
Email: isr0@cdc.gov 
 
 
Sent: Monday, November 23, 2020 10:48 AM 
To: Ananda Bandyopadhyay <Ananda.Bandyopadhyay@gatesfoundation.org>; Wilkinson, Amanda 
(CDC/DDPHSIS/CGH/GID) <lxq6@cdc.gov>; Patel, Jaymin (CDC/DDPHSIS/CGH/GID) 
<isr0@cdc.gov>; Burns, Cara C. (CDC/DDID/NCIRD/DVD) <zqd1@cdc.gov>; Jillian Gauld 
<jgauld@idmod.org>; Mike Famulare <mfamulare@idmod.org> 
Cc: Dr. K. Zaman <kzaman@icddrb.org>; Masuma Hoque <masuma.hoque@icddrb.org>; Anand, 
Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov> 
Subject: RE: Approval of nOPV2-bOPV protocol from  
 
Thanks Amanda, 
 
Approve for submission. 
 
Best, 
Jenn 
 
From: Ananda Bandyopadhyay <Ananda.Bandyopadhyay@gatesfoundation.org>  
Sent: Monday, November 23, 2020 10:35 AM 
To: Wilkinson, Amanda (CDC/DDPHSIS/CGH/GID) <lxq6@cdc.gov>; Patel, Jaymin 
(CDC/DDPHSIS/CGH/GID) <isr0@cdc.gov>; Anstadt, Jennifer (CDC/DDID/NCIRD/DVD) 
<yrq1@cdc.gov>; Burns, Cara C. (CDC/DDID/NCIRD/DVD) <zqd1@cdc.gov>; Jillian Gauld 
<jgauld@idmod.org>; Mike Famulare <mfamulare@idmod.org> 
Cc: Dr. K. Zaman <kzaman@icddrb.org>; Masuma Hoque <masuma.hoque@icddrb.org>; Anand, 
Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov> 
Subject: RE: Approval of nOPV2-bOPV protocol from  
 
Approve for submission. 
 
Best wishes, 
Ananda  
 
From: Jillian Gauld <jgauld@idmod.org>  
Sent: Thursday, November 26, 2020 11:30 AM 
To: Wilkinson, Amanda (CDC/DDPHSIS/CGH/GID) <lxq6@cdc.gov>; Anstadt, Jennifer 
(CDC/DDID/NCIRD/DVD) <yrq1@cdc.gov>; Ananda Bandyopadhyay 
<Ananda.Bandyopadhyay@gatesfoundation.org>; Patel, Jaymin (CDC/DDPHSIS/CGH/GID) 
<isr0@cdc.gov>; Burns, Cara C. (CDC/DDID/NCIRD/DVD) <zqd1@cdc.gov>; Mike Famulare 
<mfamulare@idmod.org> 
Cc: Dr. K. Zaman <kzaman@icddrb.org>; Masuma Hoque <masuma.hoque@icddrb.org>; Anand, 
Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov> 
Subject: RE: Approval of nOPV2-bOPV protocol from  

 
130 | P a g e       nOPV2 and bOPV Study             Version: 1.06                Date: 03-Dec-2020 
 
 
 
Hi Amanda, 
 
Apologies for catching this in the 11th hour, but can we add the following clarification (bolded) to the 
power calculation simulation statement on page 32? 
 
“Using our nOPV2 simulation model, we estimate prevalence and 95% CI of shedding to be 53% 
among vaccine recipients (46.4 – 59.5%) and 23% (17.8 – 28.8%) among siblings under the age of 5. 
 
Otherwise, approve for submission and happy holidays! 
 
Jillian 
 
 
From: Mike Famulare <mfamulare@idmod.org>  
Sent: Monday, November 30, 2020 9:57 AM 
To: Jillian Gauld <jgauld@idmod.org>; Wilkinson, Amanda (CDC/DDPHSIS/CGH/GID) 
<lxq6@cdc.gov>; Anstadt, Jennifer (CDC/DDID/NCIRD/DVD) <yrq1@cdc.gov>; Ananda 
Bandyopadhyay <Ananda.Bandyopadhyay@gatesfoundation.org>; Patel, Jaymin 
(CDC/DDPHSIS/CGH/GID) <isr0@cdc.gov>; Burns, Cara C. (CDC/DDID/NCIRD/DVD) 
<zqd1@cdc.gov> 
Cc: Dr. K. Zaman <kzaman@icddrb.org>; Masuma Hoque <masuma.hoque@icddrb.org>; Anand, 
Abhijeet (CDC/DDPHSIS/CGH/GID) <dvi5@cdc.gov> 
Subject: Re: Approval of nOPV2-bOPV protocol from  
 
Hi Ananda, 
 
I approve with Jillian’s correction. 
 
Mike 
 
From: Burns, Cara C. (CDC/DDID/NCIRD/DVD) <zqd1@cdc.gov>  
Sent: Thursday, December 3, 2020 12:59 PM 
To: Wilkinson, Amanda (CDC/DDPHSIS/CGH/GID) <lxq6@cdc.gov> 
Subject: RE: Approval of nOPV2-bOPV protocol from  
 
Amanda, 
I approve of the revised protocol for submission.  
 
Thanks, 
 
Cara 
 
Cara C. Burns, PhD 
Acting Branch Chief, Polio and Picornavirus Laboratory Branch 
Division of Viral Diseases 
Centers for Disease Control and Prevention 
1600 Clifton Ave. NE, MS-H17-6 
Atlanta, GA 30329