Evaluation of the Specificity of Intradermal Test With Recombinant Tuberculosis Allergen in BCG Vaccinated Healthy Volunteers
Phase: NA
Conditions: Tuberculosis
Sponsor: National Medical Research Center of Phthisiopulmonology and Infectious Diseases

 
 
STUDY PROTOCOL 
 
This document contains confidential information. 
 
ASSESSMENT OF THE SPECIFICITY OF THE INTRADERMAL TEST WITH 
TUBERCULOSIS RECOMBINANT ALLERGEN IN BCG-VACCINATED HEALTHY 
SUBJECTS 
A prospective, multicenter, open-label, cohort study of healthy adults (18 to 30 years of age) 
(asymptomatic and not at risk for tuberculosis) 
 
Protocol No. 
102/1 
Name of the investigational medicinal 
product: 
Recombinant tuberculosis allergen (Diaskintest) 
Organization responsible for the study:  
National Medical Research Center of 
Tuberculosis, Pulmonology, and Infectious 
Diseases, a federal state-financed institution of 
the Ministry of Health of the Russian Federation 
Clinical Study Leader: 
Valentina Alexandrovna Aksenova 
Anastasia Gennadyevna Samoilova 
Medical Hotline: 
+7 (495) 681-11-66 
e-mail: 
nmrc@nmrc.ru 
Person authorized to sign on behalf of the 
NMRC TPID 
Anastasia Gennadyevna Samoilova 
Initial protocol date: 
November 22, 2021 
 
 
 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
2 
 
1. 
INVESTIGATOR SIGNATURE PAGE 
INVESTIGATOR:  
I consent to:  
• 
Implement and conduct this study properly and in strict compliance with the protocol, 
GCP requirements, all applicable laws, and regulatory requirements.  
• 
Maintain the confidentiality of all information about the study and, when this information 
is submitted to a local ethics committee or other agencies, it will be filed as confidential.  
• 
Ensure that all persons involved in the study are adequately informed of the protocol, 
study intervention(s), their responsibilities, and roles in the study.  
I have fully read the protocol and agree with all its aspects.  
 
 
 
Investigator's first and last 
name 
Signature 
Date in block letters  
 
 
 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
3 
 
2. 
TABLE OF CONTENTS 
 
1. 
INVESTIGATOR SIGNATURE PAGE .................................................................................2 
2. 
TABLE OF CONTENTS ........................................................................................................3 
3. 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS .........................................5 
4. 
SIGNATURE PAGE ...............................................................................................................6 
5. 
CONTACT DETAILS .............................................................................................................7 
6. 
INTRODUCTION ...................................................................................................................9 
7. 
STUDY OBJECTIVES .........................................................................................................11 
7.1 Primary objective ...............................................................................................................11 
7.2 Secondary objectives .........................................................................................................11 
7.3 Primary endpoint ...............................................................................................................11 
7.4 Secondary endpoints ..........................................................................................................11 
7.5 Primary variables: ................................................................................................................11 
7.6 Secondary variables: ............................................................................................................11 
8. 
STUDY DESIGN ..................................................................................................................12 
8.1 Design ................................................................................................................................12 
8.2 Study population and clinical study conduct .....................................................................14 
8.3 Number of study subjects ..................................................................................................14 
8.4 Subject recruitment ............................................................................................................14 
8.5 Inclusion criteria ................................................................................................................15 
8.6 Non-inclusion criteria ........................................................................................................15 
8.7 Exclusion criteria .................................................................................................................16 
8.8 Anticipated reasons for study discontinuation...................................................................16 
8.8.1 Permanent contraindications......................................................................................16 
8.8.2 Temporary contraindications .....................................................................................16 
9. 
INVESTIGATIONAL MEDICINAL PRODUCT ................................................................17 
9.1 Composition ......................................................................................................................17 
9.2 Procedure ...........................................................................................................................17 
9.2.1 Dose and administration ............................................................................................17 
9.2.2 Packaging and labeling ..............................................................................................18 
9.2.3 Transportation of the investigational medicinal products .........................................18 
9.2.4 Storage conditions .....................................................................................................18 
9.2.5 Procedure for generating a study participant number ................................................19 
9.2.6 Blinding и unblinding of data ....................................................................................19 
9.2.7 Compliance with the administration regimen of RTA and T-SPOT.TB ...................19 
9.2.8 Accounting for the investigational medicinal products .............................................19 
9.2.9 Precautions and overdosage.......................................................................................19 
9.2.10 
Concomitant medication therapy ...........................................................................20 
9.3 Study activities ..................................................................................................................20 
9.3.1 Assessment of specificity ..........................................................................................20 
9.3.2 Safety assessments .....................................................................................................20 
9.3.3 Collection and handling of blood samples ................................................................21 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
4 
 
9.3.4 Schedule ....................................................................................................................21 
10. ETHICAL ASPECTS ............................................................................................................22 
10.1 
Remuneration of subjects ..............................................................................................22 
10.2 
Independent Safety Data Monitoring Committee ..........................................................22 
11. ADVERSE EVENTS ............................................................................................................23 
11.1 
Definitions and terms ....................................................................................................23 
11.2 
Conventional adverse events reporting .........................................................................23 
11.3 
Immediate reporting of serious adverse events by the Investigator ..............................24 
11.4 
Expedited reporting of suspected unexpected serious adverse events by the Principal 
Investigator to the LEC .............................................................................................................25 
12. DATA MANAGEMENT AND STATISTICAL ANALYSIS .............................................26 
12.1. General provisions .........................................................................................................26 
12.2 
Data management ..........................................................................................................26 
12.3 
Analyzed populations ....................................................................................................26 
12.4 
Statistical methods .........................................................................................................26 
12.5 
Sample size determination .............................................................................................27 
12.6 
Interim analysis .............................................................................................................27 
13. PROVISIONS OF THE GOOD CLINICAL PRACTICE GUIDELINES ...........................28 
13.1 
Study Participant Information and Informed Consent Procedure .................................28 
13.2 
Submission and approval of the application to the LEC ...............................................28 
13.3 
Protection of personal data of study subjects ................................................................28 
13.4 
Investigator responsibilities ...........................................................................................28 
13.5 
Curriculum vitae and registration of study staff ............................................................29 
13.6 
Training .........................................................................................................................29 
13.7 
Monitoring .....................................................................................................................29 
13.8 
Audit and inspection ......................................................................................................29 
13.9 
Definition and archiving of source data ........................................................................30 
13.10 Definition and archiving of essential documents ..........................................................30 
14. CONFIDENTIALITY OF INFORMATION AND DISCLOSURE OF INTERESTS.........31 
15. PARTNERSHIP AGREEMENT ...........................................................................................32 
16. PROTOCOL AMENDMENTS .............................................................................................33 
17. REFERENCES ......................................................................................................................34 
 
 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
5 
 
3. 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
RTA 
Recombinant tuberculosis allergen 
BCG 
Bacillus Calmette-Guérin 
WHO 
World Health Organization 
CI 
Confidence interval 
CRF 
Case report form 
IFN-γ 
Interferon-gamma 
LEC 
Local Ethics Committee 
First Volunteer In First visit of the first Volunteer 
RTA test 
Test with recombinant tuberculosis allergen  
AE 
Adverse event 
AR 
Adverse reaction 
Last Volunteer Out Last visit of the last Volunteer 
SUSAR 
Suspected unexpected serious adverse (drug) reaction 
Suspected Adverse 
Reaction 
Reporting Form 
A form of the Council of International Organizations of Medical Sciences 
Specificity 
Probability to get a negative test result in a subject without the disease 
Sensitivity 
Probability to get a positive test result in a subject with the disease  
The NMRC TPID National Medical Research Center of Tuberculosis, Pulmonology, and 
Infectious Diseases, a federal state-financed institution of the Ministry of 
Health of the Russian Federation 
CFP-10 
Recombinant culture filtrate protein 10 kDa 
ESAT-6 
Recombinant early secreted antigenic target 6 kDa 
GCP 
Good Clinical Practice 
GMP 
Good Manufacturing Practice 
ICH-GCP 
The International Conference on Harmonisation of Good Clinical Practice 
MTb 
Mycobacterium tuberculosis 
T-SPOT®.TB 
Diagnostic assay with a reagent kit for in vitro diagnosis of tuberculosis 
infection T-SPOT.TB, Oxford Immunotec Ltd., United Kingdom 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
6 
 
4. 
SIGNATURE PAGE 
Clinical Study Leader 
Valentina Alexandrovna 
Aksenova 
 
 
 
 
(date and signature) 
Clinical Study Leader 
Anastasia 
Gennadyevna 
Samoilova 
 
 
 
 
 
(date and signature) 
Principal investigator: 
Aleksandr 
Vladimirovich 
Kazakov 
 
 
 
 
 
(date and signature) 
Responsible Investigator: Yulia Yuryevna 
Kiseleva 
 
 
 
 
 
(date and signature) 
Responsible Investigator: Andrei Olegovich 
Maryandyshev 
 
 
 
 
 
(date and signature) 
Responsible Investigator: Yelena Nikolayevna 
Dolzhenko 
 
 
 
 
 
(date and signature) 
Responsible Investigator 
Anna Valentinovna 
Abramchenko 
 
 
 
 
 
(date and signature) 
Clinical Group 
Investigator 
Nadezhda Ivanovna 
Klevno 
 
 
 
 
 
(date and signature) 
Clinical Group 
Investigator 
Konstantin 
Alexandrovich Glebov  
 
 
 
 
(date and signature) 
Laboratory Group 
Investigator 
Anna Evgenievna 
Panova 
 
 
 
 
 
(date and signature) 
Laboratory Group 
Investigator 
Larisa Yurievna 
Petrova 
 
 
 
 
 
(date and signature) 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
7 
 
5. 
CONTACT DETAILS 
 
Clinical Study Leader 
Valentina Alexandrovna Aksenova, 
Doct. Med. Sci., Professor, Head of 
Pediatric and Adolescent 
Tuberculosis Department, the 
National Medical Research Center 
of Tuberculosis, Pulmonology, and 
Infectious Diseases, a federal state-
financed institution of the Ministry 
of Health of the Russian Federation 
 
Tel.: +7 (495) 681-11-66 
e-mail: nmrc@nmrc.ru 
Clinical Study Leader 
Anastasia Gennadyevna Samoilova, 
Doct. Med. Sci. 
First Associate Director of the 
NMRC TPID 
Tel.: +7 (495) 681-11-66 
e-mail: nmrc@nmrc.ru 
Principal Investigator 
Aleksandr Vladimirovich Kazakov, 
Doct. Med. Sci., senior research 
fellow of the NMRC TPID, 
tuberculosis specialist 
Tel.: +7 (495) 681-11-66 
e-mail: nmrc@nmrc.ru 
Responsible 
Investigator 
Anna Valentinovna Abramchenko, 
assistant, Tuberculosis Department, 
Faculty of General Medicine, the 
Pirogov Russian National Research 
Medical University,  
a federal state autonomous 
educational institution  
 
Tel.: +7(499)268–28-46 
e-mail: kafedra.ftiz@gmail.com 
Responsible 
Investigator 
Yulia Yuryevna Kiseleva, Cand. 
Med. Sci., Deputy Chief Physician 
for 
Medicine, Tuberculosis Center of 
Oryol Oblast, a state-financed 
healthcare institution 
Tel.: +7 (4862) 41-60-93 
e-mail: buzoptd@yandex.ru 
Responsible 
Investigator 
Andrei Olegovich Maryandyshev, 
Doct. Med. Sci., Professor, Chief 
Tuberculosis Specialist of 
Arkhangelsk Oblast and the RF 
Northwestern Federal District 
Tel.: +7(8182)-66–05-62 
e-mail: tubdisp29@gmail.com 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
8 
 
Responsible 
Investigator 
Yelena Nikolayevna Dolzhenko, 
Head of Outpatient Care, 
Clinical Tuberculosis Center of 
Ryazan Oblast, a state-financed 
institution; Chief External Pediatric 
Tuberculosis Specialist of the 
Ryazan Oblast Ministry of Health 
Tel.: +7 (4912) 24-09-13 
e-mail: rokptd@mail.ru 
Clinical Group 
Investigator 
Nadezhda Ivanovna Klevno, Doct. 
Med. Sci., Leading Research fellow 
of Pediatric and Adolescent 
Department of the NMRC TPID 
Tel.: +7 (495) 681-11-66 
e-mail: nmrc@nmrc.ru 
Clinical Group 
Investigator 
Konstantin Alexandrovich Glebov, 
Head of Outpatient Consultative 
Department, the NMRC TPID 
Tel.: +7 (495) 681-11-66 
e-mail: nmrc@nmrc.ru 
Laboratory Group 
Investigator 
Anna Evgenyevna Panova , Cand. 
Med. Sci., Head of Laboratory 
Diagnostics Department, the NMRC 
TPID 
Tel.: +7 (495) 681-11-66 
e-mail: nmrc@nmrc.ru 
Laboratory Group 
Investigator 
Larisa Yurievna Petrova, Cand. 
Med. Sci., Researcher at the Clinical 
Laboratory of Microbiology, 
Virology, and Molecular Biological 
Methods of Investigation, the 
NMRC TPID 
Tel.: +7 (495) 681-11-66 
e-mail: nmrc@nmrc.ru 
 
 
 
 
 
 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
9 
 
6. 
INTRODUCTION 
Tuberculosis remains the most common infection, affecting more than 10 million people annually 
and causing an estimated 1.2 million deaths [WHO, Global Tuberculosis report 2020]. For a 
century, tuberculin has been used worldwide to diagnose tuberculosis and detect latent tuberculosis 
infection [von Pirquet C., 1907, Mantoux M.,1912]. The major disadvantage of the tuberculin test 
is a high rate of false positive reactions due to cross-reactivity with PPD antigens that are present 
in many mycobacteria and strains of Bacillus Calmette-Guérin (BCG). In a meta-analysis of 14 
studies from other countries, the pooled sensitivity of the tuberculin test was 71% [1]. In a large 
review of foreign literature and meta-analysis, Rangaka et al. (2012) indicate that a positive 
tuberculin reaction has a low prognostic value in assessing the probability of the disease [2]. 
The limited specificity of the tuberculin skin test is particularly relevant in large-scale BCG 
vaccination programs and in the screening a large population of tuberculosis-infected patients, i.e. 
in most countries with high tuberculosis rates. The discovery of Mycobacterium tuberculosis-
specific antigens that are absent in Mycobacterium bovis BCG and most mycobacteria resulted in 
the development of in vitro tests measuring gamma interferon (INF-γ) production (Interferon-
Gamma Release Assays or IGRA) in response to stimulation with these antigens. The genome of 
Mycobacterium bovis BCG and most non-tuberculous mycobacteria has no region of difference 
(RDI) containing genes encoding these proteins. These tests showed high sensitivity and almost 
absolute specificity. 
In 2009, the diagnostic product recombinant tuberculosis allergen (RTA), a recombinant protein 
CFP10-ESAÒ6 produced by Escherichia coli BL21(DE3)/pCFP-ESAT, was granted marketing 
authorization in Russia. The Russian skin test with RTA is analogous to the IGRA-based tests. No 
special laboratory equipment is required to perform the skin test. The RTA test showed high 
sensitivity (over 95%) in the mass screening of patients in Moscow in 2012–2014 [3,4], as well as 
in Novosibirsk Oblast [5], Ryazan Oblast [6], and St. Petersburg [7].  
The following normative documentation regulates the use of the RTA test in Russia: Order of the 
Ministry of Health of the Russian Federation No. 855 of October 29, 2009; Order of the Ministry 
of Health of the Russian Federation No. 951 of December 29, 2014; Order of the Ministry of 
Health of the Russian Federation No. 124н of March 21, 2017; the clinical guidelines “Latent 
tuberculosis infection in children,” “Tuberculosis in children” (Moscow, 2020) [10], and the 
clinical guidelines “Tuberculosis in adults” (Moscow, 2020) [11].  
RTA has been used in the Russian Federation and CIS since 2009. More than 60 million tests have 
been performed since its authorization. In 2015, per Order of the Ministry of Health of Russia 
No. 951 of December 29, 2014, the RTA test replaced the Mantoux test in the screening of 
tuberculosis in children 8 years of age or older.  
Studies have shown a significant increase in tuberculosis rates (37-fold in schoolchildren aged 8–
17 years compared with the Mantoux test [12]) in primary healthcare centers following the 
transition to the screening using RTA. 
The specificity of the RTA test was also evaluated in a local (Moscow) patient population with a 
low tuberculosis rate and a low risk of tuberculosis infection (contacts of pregnant women). In 
2015–2016, 7,249 tests were performed in this population which most closely mirrors the 
permanent population of Moscow. A positive or equivocal RTA test result was found in 0.3% 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
10 
 
(95% confidence interval [CI] 0.2–0.6%) and 1.0% (95% CI 0.7–1.4%) of evaluated pregnant 
contacts (p < 0.05) in 2015 and 2016, respectively. No tuberculosis patients were identified in this 
population. Thus, the specificity of the test was 99.0–99.7% [13]. 
With decreasing incidence of tuberculosis in the Russian Federation, it seems relevant to confirm 
the specificity of the RTA test in a population of healthy subjects who are not at risk of tuberculosis 
(with a negative T-SPOT.TB assay result) in real clinical practice in the subjects of the Russian 
Federation with low rates of tuberculosis incidence and mortality. 
It should be noted that specificity, which is one of the key characteristics of a diagnostic test, is 
defined as the ability of a test to correctly identify subjects without the disease. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
11 
 
7. 
STUDY OBJECTIVES 
7.1 
Primary objective 
To evaluate the specificity of the test with recombinant tuberculosis allergen (RTA) following 
intradermal administration in healthy subjects 18 to 30 years of age, not belonging to the risk 
group for the development of tuberculosis in the regions of the Russian Federation with a 
favorable epidemiological situation for tuberculosis. 
The specificity of the RTA test is defined as the relative frequency of subjects in a healthy 
population (individuals with no signs of tuberculosis infection) who have a negative test reaction 
(no infiltration/papule) at the injection site). 
7.2 
Secondary objectives 
To record all adverse events that occurred within 28 days after the test  
7.3 
Primary endpoint 
- 
Absolute number and percentage of the volunteers with negative and positive test results 
with RTA (individuals with no signs of tuberculosis infection) based on the results of the 
assessment (measurement) of the infiltrate 72 hours after the test. 
7.4 
Secondary endpoints 
- 
Local (at the injection site) adverse events (itching, swelling, soreness, pain) within 72 
hours after administration of RTA. 
- 
General and local adverse reactions within 28 days after administration of RTA.  
 
7.5 Primary variables:  
Diameter of the infiltrate (papule) at the injection site, measured across the longitudinal axis of 
the forearm 72 hours after the intradermal test. 
7.6 Secondary variables:  
- 
Adverse events at the injection site within 28 days after the test  
- 
All adverse events that occurred within 28 days after the test  
 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
12 
 
8. 
STUDY DESIGN 
8.1 
Design 
This clinical study is a prospective, multicenter, open-label study with the formation of a cohort 
of healthy adults (18-30 years of age) (not classified as at risk of tuberculosis, with no clinical 
symptoms of the disease, with a negative T-SPOT.TB test results) to evaluate the specificity of the 
test with recombinant tuberculosis allergen following administration of the investigational 
diagnostic product to each participant at a dose of 0.2 μg/0.1 mL. Study design is presented below 
(picture 1). 
 
Picture 1. Study design 
 
In this study, in accordance with the current national standards for diagnosis of tuberculosis in 
Russia, volunteers will undergo a comprehensive examination with an assessment of clinical 
manifestations of the disease (general and respiratory complaints) and an assessment of the 
objective status. Blood sampling for the T-SPOT.TB test will be performed before the test with 
recombinant tuberculosis allergen.  
Volunteers’ complaints, as well as adverse events that occurred during the follow-up period, will 
be recorded at all visits. 
The main goal of this study is to evaluate the specificity of the skin test with recombinant 
tuberculosis allergen, defined as the relative frequency of participants in a population of healthy 
volunteers (with no signs of tuberculosis infection) whose test results are comparable to that of the 
T-SPOT.TB reference test.  
 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
13 
 
Visit 0 
Screening visit is for an initial examination, in accordance with generally 
accepted principles of diagnosis and monitoring, which includes: 
• obtaining the informed consent 
• assignment of an individual volunteer number 
• completion of the questionnaire on inclusion/non-inclusion criteria 
• collection of the volunteer’s medical history, allergic history, complaints 
• collection of information on the inclusion/non-inclusion criteria  
• physical examination, including measurement of height and weight 
• measurement of heart rate, breath rate, blood pressure 
• blood sampling for the T-SPOT.TB test. 
Visit 1 
A skin test injection visit (0-3 days post-screening) includes: 
- intradermal test with recombinant tuberculosis allergen in accordance with the 
regulatory documents in force in the Russian Federation in case of negative T-
SPOT.TB test 
- assessment of adverse events that occurred after the injection of the 
recombinant tuberculosis allergen. 
Visit 2 
An assessment visit (after 72 hours) includes: 
• assessment of the reaction to the intradermal test with recombinant 
tuberculosis allergen. 
• assessment of adverse events that occurred after the injection of the 
recombinant tuberculosis allergen. 
Visit 3 
Follow-up Visit (28 days after Visit 1):  
• assessment of adverse events that occurred after the injection of the 
recombinant tuberculosis allergen. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
14 
 
Each volunteer participating in the study will be monitored throughout the study. Anonymized 
CRFs will be shared between the study sites by courier delivery or their copies via e-mail. 
A CRF for each visit is provided in Appendix 4. 
8.2 
Study population and clinical study conduct 
Healthy individuals 18 to 30 years of age, not belonging to the risk group for the development of 
tuberculosis in the regions of the Russian Federation with a favorable epidemiological situation 
for tuberculosis.  
8.3 
Number of study subjects 
150 healthy BCG-vaccinated male and female volunteers 18 to 30 years of age. 
8.4 
Subject recruitment 
Volunteers will be recruited by the centers included in the study. They will comply with all 
inclusion criteria. Recruitment will be performed via announcements. 
The volunteer can enter screening and participate in the study only after signing the informed 
consent. After signing the informed consent the volunteer’s compliance with the inclusion / non-
inclusion criteria is assessed If a volunteer meets at least one non-inclusion criterion, he/she will 
not be able to continue his/her participation in the study. 
The T-SPOT.TB test will be performed at Visit 0 prior to the test with recombinant tuberculosis 
allergen as a reference method for diagnosis of tuberculosis infection.  
If the result of the T-SPOT.TB test is negative and other inclusion criteria are met, the volunteer 
will be enrolled in the study and can continue his/her participation in the study (the study design 
is presented in Appendix 3). 
For practical reasons, Visit 1 (product administration) should be scheduled for Monday, Tuesday, 
or Friday (to allow 72 hours follow-up after RTA administration). See section 9.3 for more 
information about visits. 
The occurrence of an infiltrate of any size at the RTA injection site will be regarded as a positive 
result. 
The T-SPOT.TB test results will be regarded as negative, indeterminate, or positive upon receiving 
results from the laboratory performing the test in accordance with the Instructions for use. If the 
result of the T-SPOT.TB test is indeterminate or positive, the volunteer will be excluded from 
further participation in the study. 
All results will be recorded in the individual Case Report Form (CRFs) of the study participants. 
Healthcare professionals will ensure that informed consent is obtained from the volunteers for the 
skin test and blood collection for research purposes. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
15 
 
8.5 
Inclusion criteria 
5. Willingness to cooperate and follow the recommendations of the Investigator in accordance 
with the Protocol. 
8.6 Non-inclusion criteria 
Subjects will not be enrolled in the study for any of the following reasons: 
1. A history of tuberculosis or close contact with a patient with active tuberculosis for 5 years 
prior to the enrollment in the study.  
2. Positive T-SPOT.TB test at the enrollment in the study 
3. Treatment with drugs affecting immune system within 3 months prior to the enrollment in 
the study 
4. Vaccination against any infections <1.5 months prior to the enrollment in the study 
5. Vaccination with BCG <6 months prior to the enrollment in the study. 
6. The Mantoux test with 2 TU and/or the test with the recombinant tuberculosis allergen was 
performed less than 6 months prior to the enrollment in the study. 
7. Congenital or acquired immunodeficiency. 
8. Active disease of the immune system  
9. HIV infection. 
10. The current condition of the skin interferes with the conduct and reading of skin tests 
(trauma, skin diseases). 
11. A disease in which blood sampling poses a risk to the volunteer (hemophilia, other bleeding 
disorders) or obstructed venous access. 
12. The volunteer currently participates in another clinical study or has participated in another 
clinical study within 3 months prior to the enrollment in the study. 
13. Previous participation in clinical studies of ESAT-6 and/or CFP-10 antigens. 
14. Pregnancy, lactation, pregnancy planning. 
15. The reluctance of a female person to use effective barrier (including spermicidal gel), 
hormonal or intrauterine contraceptives during the study. 
16. History of alcohol, drug, benzodiazepine, or other substance abuse within the 12 months 
prior to the enrollment in the study. 
17. Use of alcoholic beverages within 24 hours prior to the visit. 
18. A condition or disease that, in the opinion of the Investigator, is inappropriate for 
participation in the study. 
1. Signed informed consent for the participation in the study, as well as for the Investigator's 
access to medical records. 
2. Age 18 to 30 years 
3. A history of BCG vaccination (confirmed by medical documentation and/or the presence of 
BCG scar) 
4. Healthy individual according to physical examination and medical records at screening. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
16 
 
8.7 Exclusion criteria 
1. Refusal to participate in the study. 
8.8 
Anticipated reasons for study discontinuation 
Volunteers may discontinue participation in the study at any time without giving a reason. 
No medical events are foreseen that might cause a volunteer to drop out of the study, since invasive 
procedures are only performed during Visit 0 and Visit 1. Procedures performed at subsequent 
visits do not expose the study participant to any risk.. 
The study may be stopped at any time if the Principal Investigator concludes that participation in 
the study exposes the volunteers to unacceptable risks. 
If for any reason a volunteer wishes to withdraw from the study or has to be withdrawn from the 
study at the discretion of the Principal Investigator, the date and reason for the withdrawal should 
be documented in the CRF. 
8.8.1 Permanent contraindications 
Permanent contraindications are listed as the non-inclusion criteria in Section 8.6. 
8.8.2 Temporary contraindications 
The Responsible Investigator may, at his or her discretion, decide to postpone a visit based on the 
health assessment findings of a subject on the scheduled testing day. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
17 
 
9. 
INVESTIGATIONAL MEDICINAL PRODUCT  
One dose of the product is 0.1 mL/0.2 µg 
ATC code: V01AA20 
9.1 
Composition 
Investigational medicinal product: Recombinant tuberculosis allergen (Diaskintest).  
Recombinant protein CFP10-ESAT6 …………………………………….0.2 µg  
Sodium phosphate dibasic dihydrate ……………………..0.3876 mg  
Sodium chloride ……………………………………………………………… 0.46 mg  
Potassium phosphate monobasic………………………………. 0.063 mg  
Polysorbate 80 ……………………………………………………………… 0.005 mg  
Phenol ……………………………………………………………………….. 0.25 mg  
Water for injection ……………………………………………………... q.s. to 0.1 mL 
9.2 
Procedure  
A qualified healthcare professional will read the result of the skin test (diameter of 
hyperemia/infiltrate (papule) in mm) 72 hours after the RTA injection. An infiltrate (papule) of 
any size will be considered a positive result. 
9.2.1 Dose and administration  
The product for the skin test is administered as per the prescribing information for RTA by 
healthcare professionals who have a certificate to perform the RTA test, appropriate training, and 
experience with the test. Blood samples will be collected by a staff member who is authorized, 
trained, and experienced in blood collection for the T-SPOT.TB assay. 
The number assigned to the study subject should match the subject identification number on the 
package of the investigational medicinal product. 
Administration of the product for the skin RTA test: 
- 
The investigational medicinal product should be removed from the refrigerator half an hour 
before administration. 
- 
The expiry date should be checked before administration. 
Investigational medicinal product:  
Generic name: Standard dilution of recombinant tuberculosis allergen 
Trade name: Diaskintest  
Manufacturer: GENERIUM JSC, Russian Federation 
Dosage form: solution for intradermal injection. 
Appearance: clear, colorless liquid 
The RTA intradermal test is performed according to the current prescribing information for the 
medicinal product. An RTA dose of 0.1 mL is injected into the inner surface on the mid-third of 
the right or left forearm. The injection is performed using a 1-mL syringe with a short beveled 
needle (a tuberculin syringe).  

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
18 
 
- 
The investigational medicinal product may be administered only if it has the appearance 
of a clear and colorless solution. 
- 
Injection site disinfection is not required before administration. If the injection site does 
get disinfected, it must be allowed to dry completely before administration of the 
investigational medicinal product. 
- 
At Visit 1, the Investigator documents in which forearm the investigational medicinal 
product should be injected in the CRF.  
- 
As per the RTA prescribing information, a dose of 0.1 mL should be administered using a 
1.0-mL sterile disposable syringe with a 26-gauge short-bevel needle. 
- 
The skin is slightly stretched, and the needle is held almost parallel to the skin, with the 
bevel facing upward. The needle should be inserted about 2 mm deep into the inner surface 
on the mid-third of the left or right forearm. The needle should be visible through the 
epidermis, and 0.1 mL of the investigational medicinal product is slowly injected 
intradermally. When placed correctly, the injection should produce a small papule on the 
skin, 8 to 10 mm in diameter. The papule will resolve in about 10 minutes. 
The absence of a papule at the time of the intradermal RTA injection (as described above) should 
be documented in the CRF, providing a reason. Repeated administration of RTA to the same 
volunteer is not allowed. The volunteer should come to the follow-up visits as part of the study for 
safety monitoring. Volunteers without a papule after the intradermal injection of RTA are excluded 
from the analysis. 
After injection, the syringe and needle should be disposed of as a unit in a labeled puncture-
resistant container.  
9.2.2 Packaging and labeling 
The T-SPOT.TB assay and the RTA test are used routinely in specialized facilities, with all 
packaging requirements in place.  
9.2.3 Transportation of the investigational medicinal products 
The T-SPOT.TB assay and the RTA test are used routinely in specialized facilities. Transportation 
of the investigational medicinal product is not applicable in this study. 
9.2.4 Storage conditions 
The shelf life of RTA is 24 months when stored in a refrigerator at +2 °C to +8 °C, which will be 
monitored during the study.  
Vials should be stored in cartons until the administration of the product. The expiry date is printed 
on the outer label of the carton. 
At each study site, the investigational medicinal product should be stored in a refrigerator with a 
temperature control system and an alarm system.  
In case of malfunction of the refrigerator storing the investigational medicinal product, it should 
be placed in another refrigerator as soon as possible. The reason and time when the investigational 
medicinal product was transferred to another place should be documented in a temperature control 
log. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
19 
 
The product must not be used if the Principal Investigator has found significant deviations from 
the recommended storage conditions. The Principal Investigator should discuss with the Clinical 
Study Leader whether the product should be disposed of or can be used in the study. 
Monitoring of storage conditions shall be the responsibility of the study staff.  
9.2.5 Procedure for generating a study participant number 
On the day of screening, volunteers arriving at the center will be numbered consecutively (the first 
participant arriving will receive the lowest screening number). The individual volunteer number 
will be recorded in the informed consent and will be recorded in the individual sign-in cards at the 
visits thereafter. 
The screening number is the individual volunteer number. 
Method of generating a study participant number: 
Each volunteer will be assigned the lowest available number during visit 0.  
The available numbers in the study sites will be generated as follows: 
The first digit is the first letter of the city where the study site is located; 
The second to fourth digits are the smallest available three-digit number starting from 001. 
9.2.6 Blinding и unblinding of data 
This is an open-label study, and, hence, the study staff and subjects will be aware of the study 
stage and the products being administered. Subjects' data will only be present in informed consent 
forms which will not be shared or processed during the study, including all visits. Only study 
subject numbers will be available when the data are processed.  
9.2.7 Compliance with the administration regimen of RTA and T-SPOT.TB 
All procedures will be performed by the study staff during visits in a controlled environment. 
Monitoring of compliance with the administration regimen is not applicable in this clinical study. 
9.2.8 Accounting for the investigational medicinal products 
The T-SPOT.TB assay and the RTA test are used in routine practice. Product accounting beyond 
that is available at a given study site is not applicable.  
9.2.9 Precautions and overdosage 
As per the RTA prescribing information, transient signs of a general reaction, such as malaise, 
headache, and pyrexia, may rarely occur. These reactions, if any, will be documented in the CRF. 
No anaphylactic reactions associated with the tuberculin test have been observed. However, the 
necessary means to manage an anaphylactic reaction should always be available at the study site. 
The skin tests will be performed as per the current Russian guidelines for the diagnosis of 
tuberculosis which stipulate that these tests should be performed annually in certain 
subpopulations. 
Venous blood draws may be associated with a minimal risk which could not jeopardize life or 
health.  

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
20 
 
9.2.10 Concomitant medication therapy 
During the study, information on concomitant medication therapy considered relevant for the 
subject should be recorded on the appropriate pages of his/her CRF.  
A decision to withdraw a subject from the study will be at the discretion of the Responsible 
Investigator. Drugs and live vaccines listed in Section 8.6 are among the non-inclusion criteria. 
9.3 
Study activities 
Visit number 
0 
1 
2 
3 
Study days 
Day 
-3 
to 0 
0 
3 
28 
Informed consent procedure 
- 
assignment of a study participant number 
× 
 
 
 
Collection of the information relevant to the inclusion/non-
inclusion criteria 
× 
 
 
 
Medical history, symptoms 
× 
× 
× 
× 
Medical examination 
× 
× 
× 
 
Blood sampling for the  
T-SPOT.TB assay 
× 
 
 
 
The RTA skin test (0.2 µg) 
 
× 
 
 
Documentation of AEs 
 
× 
× 
× 
Reading of the RTA intradermal test result 
 
 
× 
 
1) A Screening Visit (Visit 0) is performed within 3 days before an Enrollment Visit (Visit 1) 
2) A Final Visit is performed if the subject discontinues the study before Visit 3 (Day 28). 
3) The medical examination which includes a review of complaints and assessment of clinical 
symptoms is performed per the current guidelines. The examination must include 
measurements of body weight, height (only at screening), and body temperature, BP, HR, RR 
(performed according to the regulations adopted in Russia). 
9.3.1 Assessment of specificity 
Specificity is a key characteristic of a diagnostic assay, referring to its ability to correctly identify 
individuals without the disease. 
The specificity of the RTA test is defined as the proportion of subjects who tested negative (no 
infiltration (papule) in the injection site) in a healthy population (i.e. without MTb exposure). 
      
 
 9.3.2 Safety assessments 
Local and systemic AEs will be evaluated based on observations made during the first hour after 
the administration of the investigational medicinal product and on health assessments performed 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
21 
 
at follow-up visits on Days 3 and 28 after the administration of the investigational medicinal 
product. Medical examinations will be performed during all visits.  
The Investigator will assess all AEs and will also report any serious AEs as described in Section 
11.3. Any local or systemic AEs should be documented in the CRF. 
All volunteers should be monitored by experienced medical staff at the study site for one hour 
after the interventions. Immediate-onset AEs, if any, should be documented in the CRF. 
The intolerance assessment criteria include the frequency of AEs that occurred during the follow-
up period. 
Note: values obtained and documented at Visit 0 will be considered baseline. 
9.3.3 Collection and handling of blood samples 
Blood samples will be collected by venipuncture performed by trained healthcare professionals as 
per the standard protocols for medical manipulations. 
Blood samples for the T-SPOT.TB assay 
The T-SPOT.TB assay is used, stored, and analyzed according to the manufacturer's manual. 
Blood is drawn from a vein into heparinized tubes (with a green cap). The first portion of blood 
should go into the heparinized tube. The volume of blood to be drawn for the study is 8 mL (one 
8-mL tube or two 4-mL tubes). Immediately after drawing of the blood sample, the tube should be 
carefully turned upside down 3–4 times to prevent foaming and without shaking so that the blood 
is evenly mixed with the anticoagulant. The tubes are transported to the laboratory in the upright 
position at a temperature between +18 ºC and +25 C in a place protected from direct sunlight. The 
tubes must not be refrigerated or frozen. Each tube must be labeled with the full name (or the 
individual number of the subject), date, and time of blood collection. The accompanying form 
must clearly and legibly state the unabbreviated full name (or the individual number of the subject), 
date of birth, time and date when the material was collected. Non-compliance to the 
recommendations for the timeframe and temperature for storage and shipping, errors in the 
accompanying form and tube labeling may result in erroneous or invalid test results.  
The T-SPOT.TB assay will be performed at certified laboratories in the participating regions. 
Subjects’ numbers will be used as identifiers to ensure their confidentiality. 
9.3.4 Schedule 
An application to authorize the study is submitted to the local ethics committee (LEC) of the 
NMRC TPID in Q4 2021.  
The date of the first visit of the first volunteer (First Volunteer In) is Q4 2021.  
The date of the last visit of the last volunteer (Last Volunteer Out) is Q1 2022. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
22 
 
10. 
ETHICAL ASPECTS 
Informed consent should be obtained from each volunteer as per local requirements after he or she 
has been informed of the nature of this study. Data collection and processing will be performed in 
compliance with all confidentiality principles. 
All study participants will be examined in accordance with normative documentation in force in 
the Russian Federation. Volunteers will be informed that their consent to participate in the study 
is voluntary and that they may withdraw it at any time, without stating the reason or being 
sanctioned.  
Approval will be obtained from the LEC of the NMRC TPID before the study commencement. 
 
The total amount of blood to be drawn from each volunteer is approximately 8 mL. Blood samples 
will be coded before being sent to the laboratory so that only the Principal Investigator and study 
staff have access to the information linking the laboratory results to the individual volunteer 
number. 
Volunteers should not expect any benefit from participating in the study, except for the benefit of 
a detailed health assessment to be conducted during the study. 
10.1 
Remuneration of subjects 
Volunteers will not be paid for participation in the study.  
10.2 
Independent Safety Data Monitoring Committee 
Subjects are expected to be exposed to only minimal risks during the clinical study. The Principal 
Investigator and Responsible Investigator of the study site shall be responsible for the safety of 
subjects in daily clinical practice. A singe fixed dose of the investigational medicinal product RTA 
will be administered to each subject during the study. Because the study has the design described 
above, an independent safety data monitoring committee is not required. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
23 
 
11. 
ADVERSE EVENTS 
This section discusses the procedures for documenting and reporting AEs during the study. 
Relevant definitions and terms are presented. Serious AEs should be immediately reported to the 
NMRC TPID.  
11.1 
Definitions and terms 
An adverse event (AE) is any unfavorable medical event detected in a patient or study participant 
after intradermal administration of the diagnostic test product and blood sampling, which may not 
have a causal relationship with its use. Thus, an adverse event (AE) can be any unfavorable 
symptom (including abnormal laboratory findings), complaint, or disease, the occurrence of which 
does not exclude a causal relationship with the use of the medicinal (investigational) product, 
regardless of the causal relationship with the medicinal product [ICH GCP E6(R2)]. 
A serious AE (experience) or reaction is any adverse medical event that is, regardless of the 
dose: 
* results in death, 
* is life-threatening, 
• Adverse events will be reported in accordance with local regulations. 
 
11.2 
Conventional adverse events reporting 
The Investigator shall be responsible for documenting in CRFs all AEs that were reported at each 
visit. The Investigator should use the following terms: 
The causal relationship between an AE and the study tests is assessed using the following terms: 
- 
Unrelated 
- 
Possibly related 
- 
Probably related 
- 
Definitely related 
The severity of an AE is assessed using the following terms: 
NOTE: the term “life-threatening” in the definition of “serious” means an event during which the 
volunteer was under threat of death; it does not concern an event that could theoretically lead to 
death, was it more severe. 
 
* requires hospitalization or prolongation of hospitalization, 
 
* leads to persistent or significant incapacity/disability or 
 
* is a congenital abnormality/malformation. 
 
• Adverse events should be recorded in the CRF using medical terminology. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
24 
 
- 
Mild (i.e. tolerated easily) 
- 
Moderate (i.e. interfering with day-to-day activities) 
- 
Severe (i.e. precluding normal functioning) 
The outcome of an AE is assessed using the following terms: 
- 
Death 
- 
Unresolved to date 
- 
Resolved with sequelae 
- 
Resolved without sequelae 
- 
Unknown 
Note. If an AE persists at the last visit, the Investigator should monitor it until it resolves or 
stabilizes. 
The seriousness of an AE is assessed based on answers to the following questions: 
- 
Did the event lead to death? 
- 
Was the event life-threatening? 
- 
Did the event require inpatient admission or prolongation of existing hospitalization? 
- 
Did the event result in severe or permanent disability or incapacity? 
- 
Does the event appear to be medically relevant? 
To avoid confusion or misunderstanding of the difference between the terms “serious” and 
“severe,” which are not synonymous, the following explanatory note is provided: 
The term “severe” is often used to describe the intensity (severity) of a specific event (e.g. mild, 
moderate, or severe myocardial infarction). However, the event itself may have relatively little 
medical significance (e.g., severe headache). The meaning of this definition is different from that 
of the term “serious,” which is based on patient/event outcomes or action criteria typically 
associated with events that are life-threatening or jeopardize the functioning of the subject. 
Seriousness (rather than severity) serves as a benchmark for determining regulatory reporting 
requirements. 
11.3 
Immediate reporting of serious adverse events by the Investigator  
The Investigator documents a serious AE (see definitions in Section 11.1) in the CRF and is 
responsible for completing the serious AE reporting form (see a template in Appendix 5). 
Any serious clinical AE (including death and overdose) that occurred during the study, regardless 
of the manipulations and procedures received by the volunteer, should be reported to the NMRC 
TPID within one working day after the serious AE occurred. 
 
The completed Suspicious Adverse Reaction Reporting Form (Appendix 5) should be sent by 
confidential mail to the following address of the Ministry of Health of Russia: ul. Dostoevskogo 
4, korp. 2, 127473 Moscow. 
 
If important additional information is discovered after the initial report, subsequent reports should 
be sent (using the same form). 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
25 
 
 
11.4 
Expedited reporting of suspected unexpected serious adverse events by the Principal 
Investigator to the LEC  
Deadlines for notifying the LEC of a suspected unexpected serious adverse reaction (SUSAR) 
after the initial information about the SUSAR was received: 
- 
For a SUSAR that resulted in death or a life-threatening condition, a notification should be 
submitted within seven (7) calendar days after the SUSAR became known to the NMRC 
TPID; the report should be submitted in additional eight (8) calendar days 
- 
For other SUSARs that are reported within fifteen (15) calendar days after the SUSAR 
became known to the LEC (prior notification is not required) 
The Safety Report Form for reporting a suspected adverse reaction must be used in SUSAR 
reports. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
26 
 
12. 
DATA MANAGEMENT AND STATISTICAL ANALYSIS 
12.1. General provisions 
Data on the proportion of subjects who tested negative in a population of healthy subjects who are 
not at risk of tuberculosis are of key importance in the development of the RTA test. 
The obtained results will allow us to conclude about the significance of the RTA test in the 
detection of tuberculosis infection. 
The NMRC TPID shall be responsible for the management of the clinical study and statistical 
analysis of data received from all study sites. 
12.2 
Data management 
Clinical data are collected in the CRF. The original CRFs remain in the custody of the study site. 
The NMRC TPID will receive copies of CRFs completed at each visit from the Responsible 
Investigators. 
The Investigator shall be responsible for proper documentation of data in the CRF of each subject 
and related records. The Investigator signing the protocol signature page shall personally sign the 
CRF to ensure that observations and findings are correctly and completely documented in the CRF. 
Copies of CRFs should be submitted to the NMRC TPID on time and will be kept by the Principal 
Investigator in a designated storage area. Investigators will be requested to clarify any ambiguous 
or implausible data if any.  
12.3 
Analyzed populations 
The population for the safety analysis includes all participants who had one RTA test during the 
study. 
The efficacy population will be equivalent to the per-protocol population and will comprise all 
participants who have met the inclusion criteria. 
12.4 
Statistical methods 
The study involves the analysis of anonymized CRF data that will be performed using an MS Excel 
spreadsheet:  
• 
Study subject number; 
• 
Enrollment date; 
• 
Sex;  
• 
Date of birth;  
• 
BCG vaccination status (presence of a scar); 
• 
Height; 
• 
Body weight; 
• 
Complaints; 
• 
Body temperature; 
• 
RR per minute; 
• 
HR; 
• 
BP (mmHg); 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
27 
 
• 
Lung auscultation: rhonchi or crackles; 
 
• 
The RTA test result (mm); 
• 
The T-SPOT.TB assay result. 
12.5 
Sample size determination 
For N patients, the variance of the estimated specificity is calculated as Var (û) = u (1–u)/N, where 
u is (unknown) the true specificity. For a fixed value N Var (û) reaches its maximum at u = 0.5. 
With N = 150, this maximum is 0.25/N = 0.0017, which corresponds to standard deviation (SD) 
(û) = √(Var (û) ) = 0.041. Since true specificity is expected to be higher than 0.5, an SD of 0.041 
would be the worst-case scenario. With a more realistic value of true specificity of 0.9 (specificity 
in studies to assess the specificity of RTA was >90%), the corresponding SD would be 0.024 which 
is considered an acceptable estimated accuracy. 
12.6 
Interim analysis 
No interim statistical analysis is anticipated. 
Generally accepted statistical methods will be employed to analyze the study data using 
Microsoft® Excel® 2016 MSO application software package (version 2109 16.0.14430.20154, 
32-bit, product code: 00333-59091-97459-AA884). The use of the software is planned for 
statistical analysis. Two-sided statistical tests at a statistical significance level of p < 0.05 will be 
used. 
Demographic and other baseline characteristics will be presented for all patients enrolled in the 
study (the full analysis set population).  
The Shapiro-Wilk criterion will be used to determine the normality of the distribution. Parameters 
that are quantitative variables (including age) will be expressed as: 
• 
Arithmetic mean (mean); 
• 
95% CI for the mean; 
Qualitative and categorical variables (gender, etc.) will be expressed in absolute values (n/N) and 
proportions (%).  
The study will evaluate the specificity of the RTA test (see. Paragraph 9.3.1). 
The sample size to assess the test specificity was initially estimated as the relative frequency in a 
binomial distribution.  

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
28 
 
13. 
PROVISIONS OF THE GOOD CLINICAL PRACTICE GUIDELINES 
This protocol has been written in compliance with the ICH guidelines, Section E6 of the ICH and 
Section E2A of the ICH, the EU directives 2001/20/EC and 2005/28/EC of the European 
Parliament (including the relevant guidance), and the National Standard of the Russian Federation 
GOST R 52379-2005 “Good Clinical Practice.” 
The study will be conducted in compliance with the principles of the Declaration of Helsinki 
(Appendix 6). 
13.1 
Study Participant Information and Informed Consent Procedure 
The subjects are informed about the study verbally and in writing, with clear explanations 
regarding the procedures, potential risks, and benefits of the study. The subjects will be given 
sufficient time (at least 24 hours) to decide whether to participate in the study.  
The informed consent must be signed and dated by the subject. A volunteer can be screened (Visit 
0) and participate in the study only after signing informed consent.  
The written information and informed consent form shall be approved by the relevant LEC before 
administration. 
13.2 
Submission and approval of the application to the LEC 
The application file will be submitted to the LEC of the NMRC TPID for approval. Enrollment of 
subjects will not begin until the study has been approved by this authority. 
The NMRC TPID is responsible for submitting the study application to the LEC. 
13.3 
Protection of personal data of study subjects 
The Responsible Investigator of the study site shall be responsible for assigning an individual 
number to a volunteer after he or she signed the voluntary informed consent. Screening failures 
should be documented in the Visit 0 CRF, including reasons for exclusion or non-inclusion. 
Voluntary informed consents and the Visits 0–3 CRFs must be signed by the Responsible 
Investigator of the study site to verify the accuracy of the entries. 
Only the individual participant number must be included in the CRF. A CRF should be always 
identifiable by the respective voluntary informed consent. 
Subjects should be aware of and agree to the following information: 
- 
After the end of the clinical study (i.e. after the Last Volunteer Out date), the Principal 
Investigator will be keeping the voluntary informed consent forms and Visit 0–3 CRFs for 
at least 15 years. 
- 
After the end of the clinical study, the voluntary informed consent forms and Visit 0–3 
CRFs should be available to the LEC or other regulatory authorities for at least 15 years, 
if required for the inspection. 
13.4 
Investigator responsibilities 
By signing this protocol, the Principal Investigator and the Responsible Investigator(s) of study 
sites shall bear overall responsibility to perform the clinical study in compliance with the protocol, 
GCP principles, and other relevant national normative documentation to ensure complete and 
accurate documenting of all data, including all AEs documented in CRFs, and to notify the NMRC 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
29 
 
TPID immediately about serious AEs as per the procedures described in Section 11 of this 
protocol. 
13.5 
Curriculum vitae and registration of study staff 
The NMRC TPID must have up-to-date, dated, and signed Russian versions of CVs of the Clinical 
Study Leader, Responsible Investigators, etc., involved in the study. 
13.6 
Training 
The Principal Investigator shall bear overall responsibility to ensure that the study staff in all study 
sites has appropriate qualifications before the study commencement. 
The Clinical Study Leader shall bear responsibility to provide training courses on general GCP 
issues, such as safety procedures, reporting of suspected serious adverse reactions including 
SUSARs, informing subjects, creating CRF entries, handling of diagnostic investigational 
medicinal products, randomization procedures, blinding and unblinding procedures, etc. If 
necessary, Responsible Investigators of the study site provide training sessions on the RTA test 
technique, etc.  
13.7 
Monitoring 
The Responsible Investigator from the NMRC TPID will oversee the conduct of the clinical study 
in the study sites. Scope of inspection includes: 
- 
Compliance with protocol requirements 
- 
Appropriate resources, equipment, and staff 
- 
Accuracy of the completion of CRFs 
- 
Concordance between the CRF and source data 
- 
Proper storage conditions of the product at the study site 
- 
Proper storage conditions of the Investigator's File at the study site 
Any issue is discussed and resolved with the Investigator or designated study staff. The oversight 
of the clinical study will be performed as per the GCP principles. 
13.8 
Audit and inspection 
The NMRC TPID has ethical, legal, and scientific obligations to oversee the conduct of the study 
per the protocol and GCP standards. 
The NMRC TPID may perform an audit of the study site. The audit will include monitoring of the 
administration of products, checking the availability of necessary documents, checking the 
procedure of signing the Information Sheet of the Informed Consent Form, and checking the 
accuracy of the completion of CRFs. 
The date and time of the audit shall be agreed upon with the Investigator involved. 
The Investigator agrees that the NMRC TPID will monitor the proper conduct and documentation 
of the study during routine visits. Data irrelevant to the study should be deleted or made 
unrecognizable. 
The Investigator consents to an audit of the study site by an auditor designated by the NMRC 
TPID, as well as by a representative of the relevant authorities and ethics committees to confirm 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
30 
 
that the conduct and documentation of the study comply with the study protocol, ICH-GCP 
regulations, and applicable law. 
The Auditor must analyze all essential documents and verify that a randomly selected sample of 
data entered in the CRFs concurs with source/primary data of clinical charts. Thus, the Investigator 
grants direct access to source data/documents and premises where the study-related procedures are 
performed. It may be necessary that the Investigator be present during the monitoring and audit 
procedures for several hours. 
13.9 
Definition and archiving of source data 
Source data are defined as all information in the original records and validated copies of the 
original records required to reproduce and evaluate a clinical study. The original documents are 
kept at the study sites. Examples of source data: 
- 
Documents approved by the LEC 
- 
Signed informed consent forms 
- 
Printed results of laboratory testing 
- 
CRFs (data entered directly into the CRF) 
- 
Consent of the study staff to participate in the study 
Copies of anonymized CRFs will be sent to the NMRC TPID by courier delivery or e-mail. 
13.10 Definition and archiving of essential documents 
The Principal Investigator is responsible for keeping essential documents in accordance with the 
ICH guidelines for at least 15 years after the study completion. The end of this clinical study is 
defined as the Last Subject Out. 
During the monitoring of the study site, the Responsible Investigator shall agree on and ensure 
compliance with the archiving requirements to complete the study. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
31 
 
14. 
CONFIDENTIALITY OF INFORMATION AND DISCLOSURE OF INTERESTS 
All CRFs, information, and results obtained by the NMRC TPID, as well as other information, 
including applications for not previously published patents and manufacturing processes, are 
considered confidential and shall remain the exclusive property of the NMRC TPID. 
An integrated statistical and clinical study report will be prepared by the NMRC TPID.  
Without a written NMRC TPID approval, no clinical study data may be published, submitted, or 
shared, except to regulatory agencies or the LEC, before the release of an internal clinical study 
report. 
All Investigators agree to avoid discussing third-party issues or publish any results of the study 
unless the NMRC TPID can provide comments. 
In the case of publication of the study results in a scientific journal, the names and order of 
appearance of authors will be determined by the NMRC TPID. 
 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
32 
 
15. 
PARTNERSHIP AGREEMENT 
The partnership agreement between the NMRC TPID and each study site should be signed before 
the enrollment of the first subject in the clinical study and should clearly define the rights and 
obligations of the parties involved.  

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
33 
 
16. 
PROTOCOL AMENDMENTS  
Clinical study procedures may be changed upon agreement with the Principal Investigator and the 
Clinical Study Leader. If the changes are substantive, the LEC approval should be obtained before 
their implementation. All substantive changes should be documented as protocol amendments, and 
a newer protocol version should be compiled as necessary. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
34 
 
17. 
REFERENCES 
1. 
 Menzies D., Pai M., Comstock G. Meta-analysis: new tests for the diagnosis of latent 
tuberculosis infection: areas of uncertainty and recommendations for research. //Ann. Intern. 
Med. - 2007. - Vol. 146. - Р. 340–354.  
2. 
 Rangaka M., Wilkinson K., Glynn J. et al. Predictive value of interferon-γ release assays 
for incident active tuberculosis: a systematic review and meta-analysis.//Lancet. Infect. Dis. 
- 2012. - Vol. 12. - Р. 45 - 55. 
3. 
 Слогоцкая Л.В., Сенчихина О.Ю., Богородская Е.М. Чувствительность теста с 
аллергеном туберкулезным рекомбинантным, содержащим белок CFP10-ESAT6, у 
впервые выявленных больных туберкулезом детей и подростков в городе Москве. 
//Туберкулез и социально-значимые заболевания. – М. – 2013. – № 1. – с. 37-44. 
4. 
 Слогоцкая Л.В., Сенчихина О.Ю., Никитина Г.В., Богородская Е.М. Эффективность 
кожного теста с аллергеном туберкулезным рекомбинантным при выявлении 
туберкулеза у детей и подростков Москвы в 2013 г. // Педиатрическая фармакология. 
- 2015. - № 1.- с. 99-103. 
5. 
 Поддубная Л.В., Шилова Е.П., Степченко И.М., Кононенко В.Г. Эпидемиологические 
факторы и иммунологические пробы в формировании групп риска по заболеванию 
туберкулезом. //Туберкулёз и болезни лёгких. - 2015. - № 5. -c. 153-154 
6. 
 Долженко Е.Н., Шейкис Е.Г., Серегина И. В. Диагностические возможности 
аллергена туберкулезного рекомбинантного в скрининг-диагностике туберкулезной 
инфекции у детей подросткового возраста в Рязанской области. // Туберкулёз и 
болезни лёгких. - 2015. - № 6. - с. 56-57. 
7. 
 Корнева Н.В., Старшинова А.А., Овчинникова Ю.Э., Довгалюк И.Ф. Сравнение 
результатов пробы Манту с 2 ТЕ и Диаскинтеста при различных проявлениях 
туберкулезной инфекции. // Туберкулез и болезни легких. – 2013. – №6. – c. 49 – 50. 
8. 
 Слогоцкая, Л. В., Филиппов А. В., Кочетков Я. А., Сельцовский П.П., Литвинов В.И. 
Чувствительность и специфичность Диаскинтеста при внелегочной локализации у 
больных с ВИЧ-инфекцией и без нее; «Иммунология», 2011, № 3, с. 116–119. 
9. 
 Рубрикатор Минздрава Клинические рекомендации «Латентная туберкулезная 
инфекция (ЛТИ) у детей», 2016 (http://cr.rosminzdrav.ru/schema.html?id=358#/text) 
10. 
 https://cr.minzdrav.gov.ru/recomend/507_1 
11. 
 https://cr.minzdrav.gov.ru/recomend/16_1 
12. 
Аксенова В.А., Моисеева Н.Н., Клевно Н.И., Одинец В.С., Баронова О.Д. 
Эффективность различных скрининговых методов раннего выявления туберкулеза у 
детей и подростков // Вопросы практической педиатрии, 2016, том 11, №4, с. 9-17 
DOI:10.20953/1817-7646-2016-4-9-17 
13. 
Богородская Е.М., Слогоцкая Л.В., Белиловский Е.М., Рощупкина О.М. Латентная 
туберкулезная инфекция в группах риска у взрослого населения города Москвы, 
2012–2016 гг. // Туберкулез и социально-значимые болезни, 2017, №2. С.10-15Pai 
Madhukar, Riley Lee W, Colford John M Jr. Interferon-D assays in the immunodiagnosis of 
tuberculosis: a systemic review. The Lancet, Infectious Diseases Vol 4 December 2004. 

Protocol No. 102/1 
November,22, 2021 
Confidential  
Version 1.1 
 
35 
 
14. 
Национальный стандарт РФ ГОСТ Р 52379-2005 «Надлежащая клиническая 
практика».