SARS-CoV-2 Infection Among Healthcare Professionals: Demographic Characteristics and Serological and Immune Responses Related to Progression's Phenotype (ProHEPiC-19)
Conditions: COVID-19, SARS-CoV-2
Sponsor: Fundacio d'Investigacio en Atencio Primaria Jordi Gol i Gurina

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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Study Protocol 
 
 
 
 
 
Title: SARS-CoV-2 infection among health care 
professionals: demographic characteristics and serological 
and immune responses related to progression's 
phenotype (ProHEPiC-19) 
 
PI and Study coordinators: Violán-Fors, Concepción; Torán-Monserrat Pere;  
Co_PIs: Garcia-Prado, Julia; Martínez-Cáceres, Eva 
 
Steering Committee: Violán Fors, Concepción; Torán-Monserrat Pere; 
Garcia-Prado; Julia; Martínez-Cáceres, Eva 
. 
 
Clinical trials protocol  id: 4r20-105  
March, 18 2021 
 
 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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INDEX 
1. HEADER ..................................................................................................................................................... 4 
2. ABSTRACT ................................................................................................................................................. 5 
3. STUDY GLOSSARY: .................................................................................................................................... 7 
4. BACKGROUND .......................................................................................................................................... 8 
5. HYPOTHESES  AND OBJECTIVES .............................................................................................................. 10 
5.1 HYPOTHESES ..................................................................................................................................... 10 
5.2 OBJECTIVES ....................................................................................................................................... 10 
5.2.1  PRIMARY  AIM........................................................................................................................... 10 
5.2.2 SECONDARY AIM ....................................................................................................................... 11 
6. METHODS ............................................................................................................................................... 11 
6.1. DESIGN: ............................................................................................................................................ 11 
6.2. POPULATION AND SETTING ............................................................................................................. 11 
6.2.1. SELECTION CRITERIA: ................................................................................................................ 11 
6.3. STUDY PERIOD ................................................................................................................................. 11 
6.4. SAMPLE SIZE (POWER) ..................................................................................................................... 12 
6.5. PROCEDURES ................................................................................................................................... 12 
6.5.1. RECRUITMENT .......................................................................................................................... 13 
6.6 VARIABLES MAIN COVARIABLES ....................................................................................................... 14 
6.6.2. OUTCOME MEASURES .............................................................................................................. 16 
6.7.DATA SOURCES ................................................................................................................................. 18 
6.8 STATISTICAL ANALYSIS ...................................................................................................................... 18 
6.8.1 STATISTIC TO MEET PRIMARY AIM: TO CONSOLIDATE A PROSPECTIVE COHORT OF HEALTH 
CARE WORKERS (HCW) TO GENERATE EPIDEMIOLOGICAL AND CLINICAL HIGH-QUALITY DATA. ..... 18 
6.8.2. STATISTICS TO MEET SECONDARY AIMS .................................................................................. 18 
6.9 ETHICS ............................................................................................................................................... 20 
7. REFERENCES ........................................................................................................................................... 21 
8.CONTRIBUTIONS OF AUTHORS AND AUTHOR AFFILIATIONS .................................................................. 22 
9.MODIFICATION OF THE PROTOCOL ......................................................................................................... 22 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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10.DISSEMINATION .................................................................................................................................... 22 
11. PROTOCOL AND REGISTRATION ........................................................................................................... 23 
12.DATA STORAGE AND SECURITY ............................................................................................................. 23 
13. ACKNOWLEDGEMENTS......................................................................................................................... 24 
14. COMPETING INTERESTS ........................................................................................................................ 24 
15. LANGUAGE............................................................................................................................................ 24 
16.  COMPENSATION .................................................................................................................................. 24 
17. OPEN ACCESS ........................................................................................................................................ 25 
18.PATIENT CONSENT DETAIL .................................................................................................................... 25 
19. PROVENANCE AND PEER REVIEW NOT COMMISSIONED; EXTERNALLY PEER REVIEWED. ................... 25 
20. DATA SHARING STATEMENT................................................................................................................. 25 
21. APENDICES ............................................................................................................................................ 25 
 
 
 
 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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1. HEADER 
TITLE OF THE PROPOSAL: SARS-CoV-2 infection among Health Care Workers: demographic 
characteristics and serological and immune responses related to progression's phenotype (ProHEpiC-19) 
PIs and Coordinators: Pere Torán Monserrat & Concepció Violán Fors 
Co-PIs: Julia Garcia Prado; Eva Martínez Cáceres   
Steering Committee: Violán Fors, Concepción; Torán Monserrat Pere; Garcia Prado, Julia; Martínez-
Cáceres, Eva. 
ORIGINAL PROTOCOL  (Written in English in March, 31 2020) 
Project members/ Contributions: 
Germans Trias i Pujol Research Institute (IGTP). Concepció Violán Fors, Pere Torán Monserrat,  Ester 
Lucas Varas, Aleix Pujol Gimeno 
Institut Universitari d'Investigació en Atenció Primària Jordi Gol (IDIAP Jordi Gol). Noemi Lamonja 
Vicente, Carla  Chacón Valera, Francesc Ramos Roure, Aleix Pujol Gimeno, Josep M Manresa Dominguez, 
Albert Roso Llorach, Lucia Carrasco Ribelles, Marta Soldevilla Garcia, Ester Moral Roldan, Magda 
Alemany Costa, Eva Olivares Ortega, Monica Monteagudo Zaragoza, Pilar Montero Alia, Marta Bujalance 
Devesa, Mariella Soto Espinoza, Cristina Palacios Fernandez, Rosalia Dacosta Aguayo.  
Institut de recerca de la Sida. IrsiCaixa: Julia Garcia Prado, Athina Kielpilanen , Oscar Blanch  Lombarte, 
Miguel Angel Marin Lopez, Julieta Carabelli, Ruth Peña Poderós, Esther Jimenez Moyano, Dan Ouchi 
Vernet, Eulalia Grau Segura. 
Atenció primària Metropolitana Nord de Barcelona. Institut Català de la Salut. Nuria Prat Gil, Josep Mº 
Bonet Simó, Nemesio Moreno Millán, Ester Badia Perich, Mónica Piña Rodriguez, Elena Domenech 
Graells, Eduard Moreno Gabriel, Victòria Sabaté Cintas, Mª Jose Argerich González, Assumpció Vazquez 
Duran, Alex Ortega Roca, Anna Devesa Pradells, Antonio Negrete. 
Hospital Universitari Germans Trias i Pujol. Institut Català de la Salut Eva Martínez Cáceres, Bibiana 
Quirant Sánchez, Gema Fernandez Rivas, Sonia Molinos Abos, Jaume Barallat Martinez de Osaba, 
Bibiana Quirant Sanchez, Joan Matllo Aguilar, Elena Domenech Graells, Fernando Armestar Rodriguez, 
Maria Doladé Botias.  
Projected timing : May, 30 2020- 30 Juny 2022 
Funding bodies:  Departament de Salut. Generalitat de Catalunya . Call COVID19-PoC number 
SLT16_04,l and  Ministerio de Ciencia e Innovación . Convocatoria COVID-10 2020. Number 
COV20/00660 
 
 
 

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2. ABSTRACT  
Introduction: Coronavirus Disease 2019 (COVID-19) has caused a global pandemic. Epidemiological and 
clinical inter-individual differences, symptomatology, recovery and humoral response against Severe 
Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) are key factors to better understand and 
predict the course of the pandemic. As Health Care Workers (HCWs) are caring for infected patients they 
are more susceptible to infection, which not only is critical for their own health but also because it 
results in a shortage of HCWs that seriously affects health services. Thus, maintaining the health and 
welfare of HCWs and enabling their rapid return to work is vital to overcome this crisis. The ProHEpiC-19 
cohort presents data on the immune response of HCWs infected with SARS-CoV-2. This dynamic cohort 
was started in March 2020 and still continues including participants.  
Objectives:  
Primary: To consolidate a prospective cohort of Health Care Workers (HCWs) to generate 
epidemiological and clinical high quality data. This information will be relevant to improve health 
policies and clinical COVID-19 protocols. This cohort will also be used as an ongoing platform to 
implement SARS-CoV-2 research projects with particular emphasis on incidence rate, reinfection, 
vaccines, and long term immune response. 
Secondary:  
1. To determine the kinetics of SARS-CoV-2 antibodies and cellular immune response in early, mid, and 
long periods of immunization.  
2.  To assess the relation between clinical variables and initial RT-PCR results with the interindividual 
differences in the immune response in early, mid, and long periods of immunization. 
3. To analyze differentially expressed cytokines as biomarkers of disease progression in early, mid, and 
long periods of immunization.  
Methods and analysis: Longitudinal, dynamic, prospective cohort study with a 12-month follow-up, 
which is being conducted in 4 primary-care centres and one hospital of Northern Metropolitana Nord of 
Barcelona (Spain). For now, the study consists of 1350 participants divided into 2 cohorts: 1) Healthy-
Exposed HCWs: 675 not infected by SARS-CoV-2 (RT-PCR with a negative result and negative SARS-CoV-2 
antibodies at baseline) and 2) Infected HCWs: 675 symptomatic participants (those with new persistent 
cough, temperature ≥37.5°C, anosmia, or ageusia or other compatible symptoms with COVID-19) or 
asymptomatic participants diagnosed by positive RT-PCR test and/or SARS-CoV-2 antibodies (IgM, IgG at 
baseline). Primary outcomes include: humoral and cellular immune response, quantitative antibodies to 
SARS-Cov-2, SARS-CoV-2 antibody levels related to progression phenotype, clinical spectrum of SARS-
Cov-2, symptomatology, demographics and other variables that may be predictive of immune response. 
Follow-up: baseline, 15 days, 1, 3, 6, 9 and 12 months. 
Findings to date: Current literature has shown that the immune response is maintained for a minimum 
of 2 months. Nevertheless little is known about the association between the immune response and the 
progression phenotype of COVID-19 .  
Future plans: This prospective cohort offers the possibility to study associations between immune 
response and progression phenotype according to age and gender as well as long-term immune 
response. In turn, we will be able to examine possible cumulative effects, taking into account several 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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clinical variables. The study is ongoing and we plan to extend it to increase the size of the cohort until 
2024. 
Ethics and dissemination: The study is approved by the Ethics Committee (IDIAPJGol ref 20/067-PCV 
IGTP ref COV20/00660 (PI-20-205). Dissemination will occur through presentations at National and 
International conferences and publications in international peer-reviewed journals. 
Trial registration number: ID: 4R20-105 
Keywords: SARS-CoV-2; COVID-19; Cohorts; Epidemiology, SARS-CoV-2 antibodies, Primary Health Care, 
Hospital, Health Care Workers, Immunology Response COVID-19. 
 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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3. STUDY GLOSSARY: 
Coronavirus Disease 2019  
 
 
 
 
 
COVID-19 
Severe acute respiratory syndrome coronavirus 
 
 
 
SARS-CoV-2 
Health Care Workers  
 
 
 
 
 
 
HCWs 
 
 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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4. BACKGROUND 
COVID-19, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), was declared 
pandemic in March 2020 by the WHO [1]. At present (March, 30, 2020), the virus has infected more than 
58.537 people worldwide with an associated case mortality rate of 1% to 15%, depending on the 
country and age [2].  
Particularly, COVID-19 has affected a very high percentage of Health Care Workers (HCWs). In fact, 26% 
of all individuals who tested positive for SARS-CoV-2 by PCR in Spain, where transmission of SARS-CoV-2 
has been intense, were HCWs [3]. So, HCWs are a high-risk group for developing the infection, that is 
exceptionally relevant for three reasons: 1) because of the clinical manifestations, severe in some cases, 
produced by the disease itself; 2) because as HCWs are caring for infected patients, they are more 
susceptible not only to infection, but also to infecting co-workers, patients and family members; and 3) 
because infection in this group leads to absenteeism from work, reducing the number of professionals 
available for health care. This situation results in a shortage of HCWs that seriously affects health 
services. Thus, maintaining the health and welfare of HCWs and enabling their rapid return to work is 
vital to overcome this crisis. Furthermore, as HCWs are constantly exposed to the virus, they are an 
interesting group to investigate the kinetics and antibody production in early, medium and long-term 
immunisations against SARS-CoV-2  
In fact, a major current question is if individuals who have been infected with SARS-CoV-2, whether 
symptomatic or asymptomatic, knowingly or unknowingly, develop permanent immunity to the virus. 
Answering this question is crucial for epidemiologists, policy makers, health service professionals and 
society at large. To do so, three different types of diagnostic tools can be used: 1) RNA-based tests to 
determine whether viral genes are present in nasal and throat swabs, indicating active infection (by RT-
PCR); 2) Detection of antibodies against SARS-CoV-2 in plasma or capillary blood. Antibodies can also 
indicate active infection but, more importantly, they indicate if the individual has already been infected 
and is able to develop a satisfactory immune response based on the presence of these antibodies. 3) 
Cytokine storm testing is a more exploratory diagnostic tool that can include assessment of the presence 
of proinflammatory cytokines in response to SARS-CoV-2 infection. The presence of a 'cytokine storm' 
has been linked to disease severity and progression and to a particular feature of the pathogenic 
features of SARS-CoV-2 infection [4].  
Different studies have shown that levels of antibody response differ depending on the clinical spectrum 
of SARS-CoV-1 [5]. Nevertheless, little is known regarding immune response to SARS-CoV-2 It has been 
observed that the highest antibody responses are presented by people with a more severe clinical 
spectrum, hospital admissions and intensive care units. The highest response has been shown by IgM 
and IgG against SARS-CoV-2 antigens [6,7]. Despite this, there are still few studies that have been able to 
accurately analyze the results of the immune response over a period of 12 months. However, the 
immune response to SARS-CoV-2 infection is heterogeneous, with clinical spectra ranging from 
individuals with asymptomatic infections to individuals with very severe clinical manifestations that can 
even lead to death. It is possible that some individuals will never develop sterilizing immunity following 
infection with SARS-CoV-2 or that multiple exposures will be needed for affinity maturation and 
development of long-lasting protection. All these points have to be explored and they are extremely 
important for re-vaccination criteria.  
At the current stage of the pandemic, laboratory tests measuring antibody response and seroconversion 
remain essential. Studies on indicators of inflammatory response and the prognostic value of other 
Covid-19 indicators and acute phase reactants are also needed. In addition, the identification of pro-
inflammatory biomarkers may have added value in disease prognosis and provide novel information for 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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the therapeutics of immunosuppression of the inflammatory process in the acute phase. Furthermore, 
by conducting serological studies of the population we can achieve the following objectives: 1) To obtain 
a dynamic, qualitative and quantitative study of the immune response to SARS-CoV-2; 2) To accurately 
define the infection rate of a specific affected area, which is essential to determine the mortality rate of 
the infection; 3) To identify individuals with strong antibody responses, who could be donors to 
generate convalescent serum therapies; and 4) To determine who has and who lacks immunity against 
the disease. This is crucial especially because this serological information can be used to strategically 
deploy HCW who are immune to infection, thus limiting the risk of exposure and inadvertent spread of 
the virus. 
As mentioned above, HCWs are a high-risk group for acquiring SARS-CoV-2 infection. Some of these 
professionals with symptoms or with high-risk contacts are confined and unable to return to their 
clinical activities while awaiting the results of the (RT)-PCR tests. It should be noted that all of them are 
confined to their homes for a minimum period of 8 days depending on the PCR result at 8, 14 and 28 
days. This is causing a shortage of HCWs, which is seriously affecting the health services and putting 
additional pressure on a sector already overburdened by the pandemic.  
Since the initial symptoms are very non-specific and compatible with any other respiratory virus, it is 
very difficult to screen for other diseases. It is therefore essential to identify factors associated with the 
protection of professionals against SARS-CoV-2. This is essential because the evolution of the SARS-CoV-
2 pandemic makes it likely that new epidemic waves will appear within a short period of time. To a large 
extent, the factors conditioning the evolution of the infection, the permanence of the virus and the 
immunity of infected persons and that of healthy carriers of the infection have not yet been clearly 
identified. 
To date, seroprevalence studies conducted in Spain have used cross-sectional designs. Thus, they cannot 
provide systematic data on long-term antibody and cellular immune responses.  
Thus, thorough serological and prospective studies are needed to determine whether ADE manifests 
among SARS-CoV-2 infections, either because of prior homologous infection or cross-reactive antibodies 
from other HCoVs. This will have particular relevance for vaccination plans. 
Another interesting point that deserves further analysis is analyzing the different antibody specificities 
generated and their dynamics. Several studies have examined various types of antibodies identified 
against SARS-Cov-2 (Spike1, Spike2, Nucleocapside, envelopes), identifying variable responses [8,9]. 
However, little is known about the simultaneous kinetics of the different antibodies over time. This can 
be analyzed with our cohort and also with the antigens used in the various vaccines that will be in the 
market in the next months. Thus, given the virus's short evolution, it is necessary to create an 
exhaustively monitored cohort. This cohort will make it possible to identify the kinetics of SARS-CoV-2-
specific antibodies in individuals with different degrees of severity of COVID-19 and to know which 
levels of antibodies are protective against new possible reinfections in 3 years. 
Crucially, this serological information will be used to strategically deploy HCW immune to the infection, 
limiting the risk of exposure and inadvertent propagation of the virus. 
In conclusion, the present study is crucial to obtain epidemiological and clinical data to improve health 
policies for the prevention of SARS-CoV-2 infection. In addition, the study will be conducted 
simultaneously with the course of the pandemic, which will make it possible to obtain real-time data on 
the different waves that occur and, in turn, to study the SARS-CoV-2 endemic. Therefore, ProHEpIC-19 
can be used as an ongoing platform to implement SARS-CoV-2 research projects with particular 
emphasis on incidence rate, reinfection, vaccines, and long term immune response.  

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5. HYPOTHESES  AND OBJECTIVES 
5.1 HYPOTHESES 
The specific hypotheses are: 
Infection with SARS-CoV-2 will produce different levels of immune response with respect to:  
1. The kinetics and immune response is different depending on the clinical spectrum of COVID-19 
disease. 
2. The immune response is conditioned by age, sex and other clinical variables  
3. Cytokines are good biomarkers to identify COVID-19 disease progression. 
5.2 OBJECTIVES 
5.2.1  PRIMARY  AIM 
To consolidate a prospective cohort of Health Care Workers to generate epidemiological and clinical 
high quality data. This information will be relevant to improve health policies and clinical COVID-19 
protocols. This cohort will also be used as an ongoing platform to implement SARS-CoV-2 research 
projects with particular emphasis on incidence rate, reinfection, vaccines, and long term immune 
response.  
Primary Outcome Measure: 
1. 
Creation prospective cohort of health care workers 
Include 675 exposed HCW participants and 675 infected HCW participants againts SARS-CoV-2, cohorts 
will be compared at each time point in terms of sociodemographic, epidemiological, clinical, and 
immunological information available. an exploratory bivariate analysis will be performed using the tests 
of Chi Square, ANOVA, Kruskall-Walis, depending on the application conditions assumptions. 
Time Frame: Baseline, to 12 months after the beginning of the study 
2. Cohort description demografics ( age, sex, academic level, housing characteristics, work variables ) 
Descriptive analysis of the participants will be performed using the number and percentage for 
categorical variables, and mean and standard deviation or median and quartiles 1 and 3 for quantitative 
variables, an exploratory bivariate analysis will be performed using the tests of Chi Square, ANOVA, 
Kruskall-Walis, depending on the application conditions assumptions. 
Time Frame: Baseline, to 12 months after the beginning of the study 
2. 
Cohort description clinical spectrum (asymptomatic, mild-moderate Illness, severe-critical) 
Cohort comparison, an exploratory bivariate analysis will be performed using the tests of Chi Square, 
ANOVA, Kruskall-Walis, depending on the application conditions assumptions. 
[Time Frame: Baseline, to 12 months after the beginning of the study] 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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5.2.2 SECONDARY AIM  
 
1. To determine the kinetics of SARS-CoV-2 antibodies and cellular immune response in early, mid, and 
long periods of immunization.  
2.  To assess the relation between clinical variables and initial RT-PCR results with the interindividual 
differences in the immune response in early, mid, and long periods of immunization. 
3. To analyze differentially expressed cytokines as biomarkers of disease progression in early, mid, and 
long periods of immunization.  
 
6. METHODS 
6.1. DESIGN:  
Dynamic prospective study conducted in two cohorts of health care workers (HCW) (healthy and 
infected). Each cohort will include 675 participants. 
Biological, clinical, psychological and social warning signs follow-up closely monitored during one year. 
6.2. POPULATION AND SETTING 
This is a multicenter study (public primary care centers set in Mataró, Sabadell and Santa Perpètua and 
the public third level hospital Hospital Germans Tries i Pujol located in Badalona) with HCW cohorts 
recruited from Gerència Territorial Metropolitana Nord of the Catalan Institute of Health (ICS), which 
consists of 7,776 HCW, including physicians, nurses, COVID-19 researchers, medical residents and other 
essential workers in direct contact with patients during present and future pandemic waves. 
6.2.1. SELECTION CRITERIA:  
HCWs (physicians, nurses, auxiliars, COVID-19 researchers and other essential workers in direct contact 
with patients during the first, second or third wave of COVID-19)  at risk of infection due to direct 
patient contact. Random sampling is being used for the inclusion of subjects. Different recruitment 
strategies are being applied to contact with participants such as email, phone calls.  
Inclusion criteria:  
≥ 18 years of age 
Accept to take part in the study and sign the informed consent  
To be a HCWs infected or exposed to SARS-CoV-2. 
Exclusion criteria: 
Not accepting taking part in the study and/or not to sign the informed consent  
Not to be a HCW exposed to SARS-CoV-2  
6.3. STUDY PERIOD 

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From March, 31 2020 until recruitment is finished (estimated on June, 15 2021). 
6.4. SAMPLE SIZE (POWER ) 
According to estimates on March, 23 2020 1350 people meet eligibility criteria. 
We will require 675 participants in each cohort to analyse the effects of SARS-CoV-2 on immunity. The 
sample will be representative of the study population. 
Sample Size Estimation 
In a one-way ANOVA study, sample sizes of 522, and 522 are obtained from the 2 groups whose means 
are to be compared. The total sample of 1044 subjects achieves 90% power to detect differences among 
the means versus the alternative of equal means using an F test with a 0.05 significance level. The size of 
the variation in the means is represented by the effect size η²= σm² / (σm² + σ²), which is 0.01 [10-13]  
Dropout-Inflated Sample Size 
Average 
Group 
Sample 
Size n 
Group 
Dropout 
Rate 
Sample 
Size Ni 
Dropout-
Inflated 
Enrollment 
Sample 
Size Ni' 
Expected 
Number of 
Dropouts 
Di 
522 
1-2 
10% 
522 
580 
58 
  
Total 
  
1044 
1160 
116 
 
 
 
 
 
Dropout- 
 
Average 
 
 
 
Inflated 
Expected 
Group 
 
 
 
Enrollment 
Number of 
Sample Size 
 
 
Sample Size 
Sample Size 
Dropouts 
n 
Group 
Dropout Rate 
Ni 
Ni' 
Di 
522,00 
1 - 2 
10% 
522 
580 
58 
 
Total 
 
1044 
1160 
116 
6.5. PROCEDURES  
Participants entering the study after meeting exclusion/exclusion criteria (Table 1), give written 
informed consent (informed consent and patient information sheet, see Appendix 1) l and they are 
assigned to a cohort  (Figure 1) . Cohorts are divided into:  
 
1) Healthy pre-cohort: HCW exposed to but not infected with SARS-CoV-2, negative PCR-test or Ig M (N) 
and IgG (N) 
2) Non-recent Infected cohort: HCW infected with SARS-CoV-2 (>15 days RT-PCR, positive)  
3) Recent Infected cohort infected HCW : HCW with acute SARS-CoV-2 infection (<15 days RT-PCR, 
positive)  
 
All participants undergo a standardised follow-up according to the specific cohort to which they are 
assigned (Table 2). At study entry, we record the following information: baseline socio-demographic 

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data, clinical data (symptoms), epidemiological data (contact with a positive and symptoms of COVID-
19) (table 3), information on the work environment, data on the diagnosis of COVID-19 (previous 
biological samples taken and dates of sick leave due to COVID-19), use of personal protective 
equipment, previous health status and habits. At subsequent visits, epidemiological data, clinical data 
(symptoms) and vaccinations are recorded.  
Finally, initial participant’s cohort assignation  is  reviewed and reclassificated on basis on laboratory 
results (RT-PCR test or SARS-CoV-2 antibodies IgM and IgG against the nucleocapsid (N)) in two final 
cohorts: 
1) Health Care Workers Healthy Cohort (HCW_H): This cohort includes HCW with negative laboratory 
test RT-PCR and SARS-CoV-2 antibodies IgM(N), and IgG(N). 
2) Health Care Workers Infected Cohort (HCW_I): This cohort includes HCW with positive laboratory test 
(RT-PCR and/or SARS-CoV-2 antibodies IgM and IgG) This cohort is subdivided into three categories 
according to the illness severity: 
Asymptomatic or Presymptomatic Infection: Individuals with positive laboratory evidence of COVID-19 
but no COVID-19 signs nor symptoms. 
Mild-Moderate Illness: Individuals with positive laboratory evidence of COVID-19 who have any of the 
various signs and symptoms of COVID-19 (e.g., fever, cough, sore throat, malaise, headache, muscle 
pain, nausea, vomiting, diarrhea, loss of taste and smell) but who do not have shortness of breath, 
dyspnea, or abnormal chest imaging and home health care.   
Severe-Critical Illness: Individuals with positive laboratory evidence of COVID-19 who have any of the 
various signs and symptoms of COVID-19 (e.g., fever, cough, sore throat, malaise, headache, muscle 
pain, nausea, vomiting, diarrhea, loss of taste and smell) and that have shortness of breath, dyspnea or 
abnormal chest imaging, treated in the hospital with or without admission to the ICU. 
This whole process is being conducted by trained multidisciplinary research staff (family physicians, 
nurses, assistant manager and project manager). See Figure 1 
6.5.1. RECRUITMENT  
The Basic Prevention Unit of Northern Metropolitan Area of Barcelona (Primary Health Care and 
Hospital), Spain (NM_BCN) offers HCW to participate in ProHEPIC-19 study, if they accept; the 
researchers of ProHEpiC-19 contact these participants. 
Eligible participants interested in the study, are accurately informed about the ProHEpiC-19 protocol, 
and signed informed consent to participate.   
Participants complete several clinical questionnaires and immune clinical determination at baseline, 7 
days, 15 days, 3, 6, 9 and 12, 16, 20, 24 months after the beginning of the study.  (see Table 2)  
Participant data files will be stored in numerical order (code system) and in a secure and accessible place 
and manner. Participant files will be maintained in storage for a period of 15 years after completion of 
the study. 
Anonymised demographic data will be collected for those participants who decline to take part in the 
project, to assess the external validity of the recruited sample of participants.  
 

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6.6 VARIABLES MAIN COVARIABLES 
1) Biological sex measured as a dichotomous variable in two categories (Male, Female) 
2) Age (years): registered and analyzed as a continuous variable 
3) Academic level: measured as a 5 levels variable (lower primary, completed primary, secondary, Post-
secondary non-tertiary education, Short-cycle tertiary education, university). 
4) Housing characteristics (meters, number of bedrooms, number of people in the same house, 
relationship, age of them) 
5) Work variables (professional category, schedule, workplace’s characteristics) 
6) Clinical Spectrum of COVID-19 defined on procedure for action against cases of infection with the new 
SARS-CoV-2 coronavirus, defined according to national and international guidelines ( asymptomatic, 
Mild-Moderate Illness, severe-critical ) as defined in section 6.5 
7) Clinical monitoring of COVID-19 symptoms, days of duration and monitoring Long Covid symptoms. 
(Table 3 , Table 4) 
8)  The date of symptom onset shall be considered as the date of appearance of the first symptoms. 
These symptoms, which are described in Table 2, are the confirmation of a positive RT-PCR test or 
antigen test (IgM (N) or IgG (N)). 
9) Diagnostics. In the electronic medical records (EMR) for Primary Care and Hospitals, the diseases are 
codified using the International Classification of Diseases, version 10 (ICD-10), including the specific 
codes for COVID-19  
10) Use of personal protective equipment (working and in the free time) 
11) Medication prescribed. Classification number of drugs according to the Anatomical Therapeutic 
Chemical (ATC) Classification System. 
12) Hospital admissions: admissions and emergency procedures according to the codes ICD-10 used in 
CMBD-HA. 
13) Comorbidities: diabetes, hypertension, ischaemic heart diseases, other chronic comorbidities 
14) Mortality by all causes. The initial data for the calculation of risk of death will be determined from 
March 15, 2020. 
15) Psychosocial and healthy habits changes during pandemic (dream, eating, sexuality, physical activity 
and familiar relationship) 
16) Laboratory Assays 
A)RT-PCR will be used as diagnostic test every 2 weeks   RNA for RT-PCR tests will be extracted from 
fresh samples using the STARMag 2019-nCoV kit (Seegene, CA, USA) on a liquid-dispensing robot 
(STARlet Hamilton, USA). To inactivate the virus and extract viral RNA, 500 μl of the transfer medium 
containing the swab is combined with 500 μl of lysis buffer of the STARMag kit in a BSL-2 laboratory; 
following 10′ of incubation at room temperature, 350 μl was used for nucleic acid extraction into a 
volume of 100 μl of extracted genome, from which 8 μl was taken for the 2019-nCoV PCR assay.The 
detection of RNA will be performed using the Allplex™ SARS-CoV-2 Assay is a multiplex real-time PCR 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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assay to detect 4 target genes of SARS-CoV-2 in a single tube. The assay is designed to detect RdRP, S 
and N genes specific for SARS-CoV-2, and E gene for all of Sarbecovirus including SARS-CoV-2 (Seegene, 
CA, USA) according to the manufacturer’s instructions. 
B) Quantiﬁcation of antibodies to SARS-CoV-2. We performed a previous validations study with 6 
different IVD-CE approved ELISA tests and selected the commercially available test kits - the AntiSARS-
CoV-2 IgG and IgM ELISA (Inmunodiagnostic). This ELISA test had a Sensitivity 92.5% and a specificity 
93.3% for IGM and IgG against Nucleocapsid (N) and Spike (S) antigens. For this, the best reagents and 
the most relevant antigen specificities will be analysed. We will perform test at baseline, 7 days, 15 days, 
3, 6, 9 and 12,16,20,24 months after the beginning of the study.  
We conducted a pre-validation study (March-April 2020) with 6 different IVD-CE approved 
ELISA tests and selected commercially available Anti-SARS-CoV-2 IgG and IgM ELISA kits 
(Inmunodiagnostic Limited ©). These ELISA tests had a sensitivity of 92.5% and a specificity of 
93.3% for IGM and IgG against the nucleocapsid (N) antigen. Positivity thresholds were 
provided by the assay manufacturers and were considered positive with an index value greater 
than 1.1, indeterminate from 0.9 to 1.1 and negative if <0.9 index units.  
Positive participants with Rt-PCR + or IgM (N) or IgG (N) were also tested for antibodies against 
the Spike (S) subunit of SARS-CoV-2. This was done by enzyme immunoassay for the qualitative 
determination of IgG class antibodies against COVID-19 (SARS-CoV-2) in human serum with 
DECOV1901.  This reagent complies with the technical specifications for sensitivity, specificity, 
and standardisation according to WHO International Standards and Reference Panel for anti-
SARS-CoV-2 antibody (WHO/BS.2020.2403). Positivity thresholds were provided by the assay 
manufacturers and were considered positive with an index value greater than 40, 
indeterminate from 32 to 40 and negative if <32 Ul/ml 
Positive participants for serology or/and RT-PCR were also tested for antibodies against the 
Spike (S) subunit of SARS-CoV-2. This was done by enzyme immunoassay for the quantitative 
determination of IgG class antibodies using DECOV1901 (Demeditec Diagnostics GmbH©).  
This reagent complies with the technical specifications for sensitivity, specificity, and 
standardisation according to WHO International Standards and Reference Panel for anti-SARS-
CoV-2 antibody (WHO/BS.2020.2403). Positivity thresholds were provided by the assay 
manufacturers and were considered positive with an index value greater than 40, 
indeterminate from 32 to 40 and negative if <32 Ul/ml 
 
C) Additional Immune response laboratory assays: 
C1) Additionally, blood samples of HCW (infected and healthy participants) will be collected and stored 
for more exhaustive determinations of the immune response in later studies. 
C2) INFg Elispot assays. To monitor both SARS-CoV-2 specific CD4+ and CD8+ T-cell responses we 
performed an IFNγ ELISPOT assay. The design compromises a comprehensive screening using peptide 
pools for the nucleocapsid, membrane, S1 and S2 regions of the spike protein described in Olvera et al., 
(2020) vaccines and total recombinant proteins for the nucleocapside and the spike. In brief, cells will be 
thawed and incubated overnight at 37°C in the presence of SARS-CoV2 peptide pools described (5ug/ml 
for each peptide) and PHA (15 μg/ml), as positive control. Finally, plates were revealed with biotinylated 
anti-human IFNγ, streptavidin-alkaline phosphatase and its coloured substrate (Mabtech, Sweeden). 

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IFNγ secreting cells were quantified under Immuno Capture and Immuno Spot software to calculate the 
number of Spot Forming Cells (SFC). Cryopreserved PBMCs will be stimulated overnight with CoV-2 
peptide pools (5ug/ml for each peptide) and PHA (15 μg/ml), as the positive control. Spot Forming Cells 
(SFC) will be counted using an INFg-ELISpot kit (Mabtech), AP-conjugate kit (BioRad) and an automated 
ELISPOT Reader Unit (CTL) as previously described (Dalmau et al AIDS 2014). Wells will be considered 
positive if they contained at least 50 spot-forming cells per 106 PBMCs above the background level (2X 
mean + 3Xstandard deviation). 
C3) Citokines multiplex assay. Cryopreserved plasma samples will be used in a 45-plex assay of soluble 
mediators. The plates will be read with a Luminex instrument (Luminex 200, Austin Luminex, USA) and 
data will be analysed using MILLIPLEX Analyst 5.1 software (Merck Millipore Darmstadt, Germany). 
Measures will be performed in duplicates. 
C4) Combinatorial flow-cytometry of lineage, activation inducible markers and intracellular cytokines 
Cryopreserved PBMCs will be stimulated with reactive peptide pools identified by ELISpot and incubated 
overnight. In the ICS, we will combine T-cell lineage markers (CD3, CD4, CD8, CD45, CCR7, CD27), 
activation inducible markers (AIMs: OX40, CD25 and CD137) and cytokines (IL2, IFNg, TNF). Samples will 
be acquired on an LSR Fortessa cytometer (BD), data analysed using FlowJo software v10 and 
polyfunctional cytokine profiling represented using SPICE v6 (NIAID, NIH).  
6.6.2. OUTCOME MEASURES 
Outcome measures of immune response (SARS-CoV-2 Antibodies (N), cytokines, T-cells) and Covid-19 
clinical symptoms will be measured at baseline and prospectively aiming to observe possible 
associations between possible prognostic factors and immune response. The baseline and prospective 
measurements will allow to assess their prognostic role and whether some of them appear as 
confounding factors. 
Primary Outcome Measure: 
1. 
Creation prospective cohort of health care workers 
Include 675 exposed HCW participants and 675 infected HCW participants againts SARS-CoV-2, cohorts 
will be compared at each time point in terms of sociodemographic, epidemiological, clinical, and 
immunological information available. an exploratory bivariate analysis will be performed using the tests 
of Chi Square, ANOVA, Kruskall-Walis, depending on the application conditions assumptions. 
[Time Frame: Baseline, to 12 months after the beginning of the study] 
2.Cohort description demografics ( age, sex, academic level, housing characteristics, work variables ) 
Descriptive analysis of the participants will be performed using the number and percentage for 
categorical variables, and mean and standard deviation or median and quartiles 1 and 3 for quantitative 
variables, an exploratory bivariate analysis will be performed using the tests of Chi Square, ANOVA, 
Kruskall-Walis, depending on the application conditions assumptions. 
[Time Frame: Baseline, to 12 months after the beginning of the study] 
2. 
Cohort description clinical spectrum (asymptomatic, mild-moderate Illness, severe-critical) 
Cohort comparison, an exploratory bivariate analysis will be performed using the tests of Chi Square, 
ANOVA, Kruskall-Walis, depending on the application conditions assumptions. 

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[Time Frame: Baseline, to 12 months after the beginning of the study] 
Secondary Outcome Measures: 
4. 
Kinetics of SARS-CoV-2. IgM Nucleocapside 
IgM (nucleocapside) ELISA kits (Inmunodiagnostic Limited ©). Positivity thresholds were provided by the 
assay manufacturers and were considered positive with an index value greater than 1.1, indeterminate 
from 0.9 to 1.1 and negative if <0.9 index units 
[Time Frame: Baseline, 7 days, 15 days, 3, 6, 9 and 12 months after the beginning of the study] 
5. 
Kinetics of SARS-CoV-2. IgG Nucleocapside 
IgG (nucleocapside) ELISA kits (Inmunodiagnostic Limited ©). Positivity thresholds were provided by the 
assay manufacturers and were considered positive with an index value greater than 1.1, indeterminate 
from 0.9 to 1.1 and negative if <0.9 index units 
[Time Frame: Baseline, 7 days, 15 days, 3, 6, 9 and 12 months after the beginning of the study] 
6. 
Kinetics of SARS-CoV-2. IgG Spike 
IgG (spike). ELISA kits DECOV1901 (Demeditec Diagnostics GmbH©). Positivity thresholds were provided 
by the assay manufacturers and were considered positive with an index value greater than 40, 
indeterminate from 32 to 40 and negative if <32 Ul/ml 
[Time Frame: Baseline, 7 days, 15 days, 3, 6, 9 and 12 months after the beginning of the study] 
7. 
Kinetics of SARS-CoV-2. T-Cell 
SARS-CoV-2 specific CD4+ and CD8+ T-cell responses we performed an IFNγ ELISPOT assay. Wells will be 
considered positive if they contained at least 50 spot-forming cells per 106 PBMCs above the 
background level (2X mean + 3Xstandard deviation). 
[Time Frame: Baseline, 7 days, 15 days, 3, 6, 9 and 12 months after the beginning of the study] 
8. 
To assess the relation between clinical variables and initial RT-PCR results in the whole sample 
and by sex. 
To study the differences between clinical spectrums and initial RT-PCR we will use ANOVAs or Kruskal-
Wallis tests, after checking normality assumption using a Shapiro-test 
[Time Frame: Baseline, to 12 months after the beginning of the study] 
9. 
To analyse the relation between clinical variables and the interindividual differences in the 
immune response in early, mid, and long periods of immunization in the whole sample and by sex 
To study the differences between clinical spectrums and immune response in early period we will use 
ANOVAs or Kruskal-Wallis tests, after checking normality assumption using a Shapiro-test . Similarly, to 
look for differences in antibody levels between sex, either a t-test or a Mann-Whitney test will be 
performed. 
[Time Frame: Baseline, 7 days, 15 days, 3, 6, 9 and 12 months after the beginning of the study] 
10. 
Cytokines as biomarkers of disease progression in early, mid, and long periods of immunization. 

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Cryopreserved plasma samples will be used in a 45-plex assay of soluble mediators. The plates will be 
read with a Luminex instrument (Luminex 200, Austin Luminex, USA).Appropriate statistical tests (i.e. t-
test or Mann-Whitney to compare between sexes and ANOVA or Kruskal-Wallis to compare between 
clinical spectrums) will be used after checking for normality (Shapiro-test) 
[Time Frame: Baseline, 7 days, 15 days, 3, 6, 9 and 12 months after the beginning of the study] 
6.7.DATA SOURCES 
We will simultaneously use various information sources 
A) Database created for the register of ProHEpiC-19. 
The data will be centralized in an internal centralized data warehouse from the Catalan Health Institute. 
After data collection and curation, the data is ready to be analyzed to (1) validate the accuracy on 
demographic, clinical, social, and psychological data of professionals; (2) register SARS-COV-2 
antibodies; (3) results in RT-PCR test. 
B) Primary care electronic health records (e-Cap) contain demographic data and clinical variables: health 
conditions (ICD-10 codes) and diagnostic date, medication prescribed, comorbidities, lab test results 
(including RT-PCR). 
C) Case Report Form (Teleform) specific for the study, which will include a specific identifier for the 
patient and the serological and immunological tests' information. This information will be shared among 
the various sources of information about the study employing individual, anonymized participants' 
identification. Immunological data will be collected to understand the vaccine's protection and the 
relationship between immune response and reinfection. 
6.8 STATISTICAL ANALYSIS 
Data collection: The data will be exported to the statistical packages used in the study.  The people that 
meet eligibility criteria will be identified and included in the study database. A comprehensive cleaning 
of the data will be conducted to eliminate inconsistencies, duplicities and treatment indications 
inconsistent with the diagnosis and analysis of missing data will also be conducted. Additionally, a 
longitudinal validation of diagnostics and medicines will be carried out. 
6.8.1 STATISTIC TO MEET PRIMARY AIM: TO CONSOLIDATE A PROSPECTIVE 
COHORT OF HEALTH CARE WORKERS (HCW) TO GENERATE EPIDEMIOLOGICAL AND 
CLINICAL HIGH-QUALITY DATA. 
A descriptive analysis of the participants will be performed using the number and percentage for 
categorical variables, and mean and standard deviation or median and quartiles 1 and 3 for quantitative 
variables. Frequency and percentage of followed-up patients and dropouts will be updated at each 
timepoint. Profiles of dropouts and visited people will be assessed using the appropriate statistical test 
in order to prevent potential biases due to drop out.  Cohorts will be compared at each time point in 
terms of sociodemographic, epidemiological, clinical, and immunological information available. To 
analyze this, an exploratory bivariate analysis will be performed using the tests of Chi Square, ANOVA, 
Kruskall-Walis, depending on the application conditions assumptions. 
6.8.2. STATISTICS TO MEET SECONDARY AIMS   

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6.8.2.1. TO DETERMINE THE KINETICS OF SARS-COV-2 ANTIBODIES AND CELLULAR 
IMMUNE RESPONSE IN EARLY, MID, AND LONG PERIODS OF IMMUNIZATION.  
Two methods will be followed to study the kinetics of SARS-CoV-2 antibodies since symptoms onset. 
Locally Stimated Scatterplot Smoothing (LOESS) models will be fitted as a first descriptive approach. 
Then, nonlinear mixed-effects (NLME) models will be fitted to get the curves that would determine the 
kinetics. 
LOCALLY ESTIMATED SCATTERPLOT SMOOTHING (LOESS) MODELS 
LOESS models will fit as an initial descriptive manner of studying the antibodies, both IgM and IgG, 
evolution since diagnosis day. LOESS regression consists of fitting simple models (e.g., first or second 
degree polynomials) in subsets of data determined by a nearest neighbours algorithm. These LOESS 
curves will best stratified either per clinical condition or gender, and will be shown together with their 
95% confidence interval (CI). We will use all the data available except for those from participants who 
never tested positive, grouping the data points per participant in the figures.  
NONLINEAR MIXED-EFFECTS (NLME) MODELS 
Mixed-effects models are generally useful when there are multiple measures per participant, as in this 
study. Random effects are included in the model to test if they may be causing any significant, and 
generally unexpected, significant influence in the modelling built by the fixed effects. Therefore, 
introducing random effects in the model help to control unobserved variance. 
We will use NLME models to study possible differences in the dynamics (kinetics) of SARS-CoV-2 
antibodies (IgM, IgG) levels since symptoms onset. After fitting a general model, the differences will be 
studied between clinical spectrums and biological sex, setting the largest group as reference groups for 
modelling. To fit these models, we will use all the data available except for those from participants who 
never tested positive. Time after diagnosis was discretized as follows: Tests made in the first 14 days 
since diagnosis are treated as "Day 15". Tests from days 15 to 29, as "Day 30", days 30 to 59 as "Day 60", 
days 60 to 89 as "Day 90", days 90 to 179 as "Day 180", days 180 to 269 as "Day 270", days 270 to 360 as 
"Day 360", and days 360 up to end of follow-up as “Day 450”. However, antibody values beyond 450 
days after diagnosis were not considered, as they were only available for 14 participants. 
The general equations to be fitted will be: 
𝐼𝑔𝑀(𝑡) = 𝑏1 + (𝑏0 −𝑏1)𝑒−𝑘1𝑡−𝑏1𝑘1(𝑒−𝑘1𝑡−𝑒−𝑘2𝑡) 
𝐼𝑔𝐺(𝑡) = 𝑏1 + (𝑏0 −𝑏1)𝑒−𝑘1𝑡−𝑏1𝑘1(𝑒−𝑘1𝑡−𝑒−𝑘2𝑡) 
Having t normalized per the maximum number of days (360). This general equation corresponds to an 
equation describing an exponential rise followed by an exponential decay. 𝑏0 corresponds to the 
baseline value, 𝑏1corresponds to the asymptotic value (i.e., antibody levels at 𝑡 →∞), 𝑘1 corresponds 
to the rise rate, and 𝑘2 to the decay rate.  
Nonlinear mixed-effects models will be built using the nlme package [14]. 
6.8.2.2. TO ASSESS THE RELATION BETWEEN CLINICAL VARIABLES AND INITIAL RT-PCR 
RESULTS WITH THE INTERINDIVIDUAL DIFFERENCES IN THE IMMUNE RESPONSE IN 
EARLY, MID, AND LONG PERIODS OF IMMUNIZATION. 
 

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The incidence by SARS-CoV-2 will be linked to the sociodemographic, epidemiological, clinical and 
immunological information collected in the project. To analyze this, an exploratory bivariate analysis will 
be performed using the appropriate bivariate tests. Chi-squared tests will be used to find if there were 
any significant differences between the expected and the observed frequencies in categorical 
comparisons (e.g., the prevalence of symptoms per sex or clinical spectrum).. To study the differences 
between clinical spectrums and initial RT-PCR we will use ANOVAs or Kruskal-Wallis tests, after checking 
normality assumption using a Shapiro-test . Post-hoc analyses will be performed to find significant 
differences between clinical spectrum and immune response. Similarly, to look for differences in 
antibody levels between sex, either a t-test or a Mann-Whitney test will be performed. 
 
6.8.2.3. TO ANALYZE DIFFERENTIALLY EXPRESSED CYTOKINES AS BIOMARKERS OF 
DISEASE PROGRESSION IN EARLY, MID, AND LONG PERIODS OF IMMUNIZATION 
Cytokine levels will be studied and compared between different clinical spectrum or sexes to determine 
their role as a biomarker of disease type. Appropriate statistical tests (i.e. t-test or Mann-Whitney to 
compare between sexes and ANOVA or Kruskal-Wallis to compare between clinical spectrums) will be 
used after checking for normality (Shapiro-test).Significance level will be set to 0.05. All the analyses will 
be performed using R version 3.6.3 or higher. 
 
6.9 ETHICS 
The biological samples for this investigation are available and stored under different IRB-approved study 
protocols and properly consented for investigational use. Study participants are informed about the 
objectives of the study and the interventions linked to their participation in it. They are also given a 
written information sheet detailing all the information about the project and asked to sign the informed 
consent form. The confidentiality and anonymity of the data are ensured in accordance with current 
state laws (Organic Law 3/2018, of 5 December, on Personal Data Protection and guarantee of digital 
rights). The study is being conducted in accordance with the articles pertaining to the Declaration of 
Helsinki, agreed at the 64th General Assembly in 2013. This project is under the tutelage of the Clinical 
Research and Ethics Committee (CEIC) of the Instituto Universitario de Investigación en Atención 
Primaria (IDIAP Jordi Gol) and incorporates its recommendations and suggestions.  
Your samples may only be used for the purpose stated in this information sheet. Your samples may be 
used in ongoing studies of COVID-19 infection, but may also be retained for future research related to 
COVID-19 infection.   
All studies using your samples must be approved by a Research Ethics Committee. You may request to 
be informed about the studies in which your samples have been used.   
Your samples will be stored mainly in the Germans Trias i Pujol Health Sciences Research Institute (IGTP) 
Biobank, an institution that has been accredited by the health authorities (Insitituto de Salud Carlos III. 
Ministerio de Ciencia e Innnovación) specifically to store samples for research under the conditions 
required by law (number B.0000643) and IrsiCaixa biological collection “Pathogenesis of infectious 
diseases” with the registration number C.0006008. The treatment and use of all samples will be carried 
out in accordance with the Biomedical Research Act (14/2007) and Royal Decree 1716/2011, which 
regulates biobanks. The samples will be kept for 15 years. They may be transferred to third parties if the 
current Biobank legislation is complied with and they will not be sold under any circumstances. 

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In some cases, they may become part of sample collections accredited by the health authorities. In this 
case, they can neither be transferred to third parties nor sold, but they could later be integrated into the 
Biobank of our centre.  
The samples will be identified in a coded form. This means that the information on the labelling of the 
samples cannot be linked to the person who donated the sample. Please see below the section on 
Confidentiality and data protection. 
The biological samples for this investigation are available and stored under IGTP and IrsiCaixa specific 
research collection-approved study protocols and properly consented for investigational use. The study 
will be approved by the Ethics Committee (IDIAPJGol ref 20/067-PCV IGTP ref COV20/00660 (PI-20-205). 
The study will be approved by the Ethics Committee (IDIAPJGol ref 20/067-PCV IGTP ref COV20/00660 
(PI-20-205) 
7. REFERENCES 
1. 
https://www.who.int/news/item/30-01-2020-statement-on-the-second-meeting-of-the-
international-health-regulations-(2005)-emergency-committee-regarding-the-outbreak-of-novel-
coronavirus-(2019-ncov) ] 
2. 
https://coronavirus.jhu.edu/us-map 
3. 
Chen ZM, Fu JF, Shu Q, Chen YH, Hua CZ, Li FB, et al. Diagnosis and treatment 
recommendations for pediatric respiratory infection caused by the 2019 novel coronavirus. World J 
Pediatr.2020, http://dx.doi.org/10.1007/s12519-020-00345-5. 
4. 
García-Sierra RM, Badia Perich E, Manresa Dominguez JM, Moreno Millan N, Sabaté Cintas V, 
Romero Martínez M, Moreno Gabriel E, Pera G, Seda Gombau G, Montellà Jordana N, Violan Fors C, 
Argerich González MJ, Bonet Simó JM, Prat Gil N, Torán Monserrat P. Descriptive study of the health 
service workers of a Primary Care Department confined by Covid-19.. Rev Esp Salud Publica. 2020 Sep 
3;94:e202009106. Spanish. PMID: 32880381. 
5. 
Liu W, Fontanet A, Zhang PH, et al. Two-year prospective study of the humoral immune 
response of patients with severe acute respiratory syndrome. J Infect Dis 2006;193:792-5. 
6. 
Callow KA, Parry HF, Sergeant M, Tyrrell DA. The time course of the immune response to 
experimental coronavirus infection of man. Epidemiol Infect 1990;105:435-46 
7. 
Choe PG, Perera RAPM, Park WB, et al. MERS-CoV Antibody Responses 1 Year after Symptom 
Onset, South Korea, 2015. Emerg Infect Dis 2017;23:1079-84. 
8. 
Huang, A.T., Garcia-Carreras, B., Hitchings, M.D.T. et al. A systematic review of antibody 
mediated immunity to coronaviruses: kinetics, correlates of protection, and association with severity. 
Nat Commun 11, 4704 (2020).  
9. 
Deeks JJ, Dinnes J, Takwoingi Y, Davenport C, Spijker R, Taylor-Phillips S, Adriano A, Beese S, 
Dretzke J, Ferrante di Ruffano L, Harris IM, Price MJ, Dittrich S, Emperador D, Hoof  L, Leeflang MMG, 
Van den Bruel A. Antibody tests for identification of current and past infection with SARS‐CoV‐2. 
Cochrane Database of Systematic Reviews 2020, Issue 6.  
10. 
Cohen, Jacob. 1988. Statistical Power Analysis for the Behavioral Sciences. Lawrence Erlbaum 
Associates. Hillsdale, New Jersey  

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11. 
Desu, M. M. and Raghavarao, D. 1990. Sample Size Methodology. Academic Press. New York.  
12. 
Fleiss, Joseph L. 1986. The Design and Analysis of Clinical Experiments. John Wiley & Sons. New 
York.  
13. 
Kirk, Roger E. 1982. Experimental Design: Procedures for the Behavioral Sciences. Brooks/Cole. 
Pacific Grove, California. 
14. 
Pinheiro J, Bates D, DebRoy S, Sarkar D, R Core Team (2020). nlme: Linear and Nonlinear Mixed 
Effects Models. R package version 3.1-144. Available at: https://CRAN.R-project.org/package=nlme 
8.CONTRIBUTIONS OF AUTHORS AND AUTHOR AFFILIATIONS  
Steering Committee: Violán Fors, Concepción; Torán Monserrat Pere; Garcia Prado, Julia;   Martínez-
Cáceres, Eva. 
PIs and Coordinators: Pere Torán Monserrat & Concepció Violán Fors 
Co-PI: Julia Garcia Prado; Eva Martínez Cáceres   
Project members/ Contributions: 
9.MODIFICATION OF THE PROTOCOL  
Changes from the original plan will be summarized on a separate paper an create a new version of study 
protocol 
10.DISSEMINATION  
Core to the achievement of the increased awareness and impact of the SARS-CoV-2 immune response is 
the improvement of communication at every level and in particular between health professionals and 
patients. 
In support of the project’s objectives, a stakeholder communications strategy will be developed, which 
links the various groups to be engaged during the project with the most effective means of providing 
them with overall messages and ongoing engagement. The strategy will include: 
a) Project visual identity 
b) Key messages 
c) Timing and content of research briefings and policy notes 
d) Schedule of public activities. 
At the same time, the project’s and social media channels will be established. The strategy will detail 
specific outputs relating to the general media. The activities detailed in the strategy will be reviewed 
every six months. 
Target groups for Dissemination and Communication 
The communication activities to the target groups are outlined in the list below. Activities will target the 
following main stakeholders: (1) HCWs; (2) Health agencies, Health politicians; (3) Patients and patient 
organization. 

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a) Social networks: Over 100 stakeholders engaged at the month 12 ProHEpiC-19 project ,  and 1000 by 
the end of the project. 
b) Briefing notes: At least six short, non-expert, briefing notes produced and distributed 
c) Press Releases: At least three, non-expert, briefing notes produced and distributed 
d) Publications and conference presentations: At least three scientific papers submitted and six 
conference presentations by the end of the project including a final conference. 
Publications and conference presentations will be targeted according to the direction of the findings and 
to take advantage of complementary conference and journal special issue topics. 
11. PROTOCOL AND REGISTRATION  
This study is registered with ClinicalTrials.gov ID: 4R20-105 
 
12.DATA STORAGE AND SECURITY 
1) Type and format of data collected/generated within the Project 
All data will be processed in a pseudo-anonymised way by coding the identifying variables in both the 
initial paper data collection booklet (CB) and the subsequently created database. Only the necessary 
data to achieve the research objectives will be collected. The necessary variables to conduct the project 
are: 1) 1) Biological sex; 2)Age (years); 3) Academic level; 4) Housing characteristics ; 5)Occupational 
situation ; 6) Clinical Spectrum of COVID-19; 7) Clinical monitoring of COVID-19 Symptoms; 8) Date  first 
symptoms; 9) Diagnostics. In the electronic medical records; 10) Use of personal protective equipment; 
11) Medication prescribed; 12) Hospital admissions; 13) Comorbidities; 14) Mortality by all causes; 15) 
Psychosocial and healthy habits changes during pandemic; 16) Laboratory Assays: RT-.PCR test, 
antibodies to SARS-CoV-2 ( IgM and Ig G against nucleocapside, IgG Against Spike) , INFg Elispot assays; 
Citokines multiplex assay; Combinatorial flow cytometry of lineage, inducible activation markers and 
intracellular cytokines. These variables can be consulted in the corresponding section of the 
methodology.   
In accordance with Law 14/2007, the samples of each participant in the project will be collected directly 
from the person concerned and the procedure set out in the regulations on the use of biological samples 
will be followed to authorise their use. The samples will only be used for the project and will 
subsequently be destroyed. 
2) Procedure provided to access data (who, how and when can access), data ownership, repository to 
deposit data. 
-The necessary variables to perform the study will be obtained directly from the participants in the 
project by means of a self-completed form to be filled in by the participant. The exploratory variables, 
biological samples and complementary explorations will be obtained by accredited personnel. All 
variables and data obtained will require the prior consent of the participant, in accordance with the 
provisions of articles 6.1.a) and 9.2.a) of the RGPD, as well as additional provision 17.2.d of the LOPD-
GDD.  

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The Catalan Institute of Health (ICS), the IDIAP Jordi Gol Research Institute and the Germans Trias i Pujol 
Research Institute (IGTP) act as data controllers in the framework of this observational study. The 
project database will be housed in the computers of the Unitat de Suport a la Recerca Metropolitana 
Nord of the ICS, which will act as data processor. Initially, international data transfer is not foreseen. If 
this were to take place, a new informed consent would be requested from the study participants.  
3) Procedure planned to ensure specific ethical and legal requirements.  
The database has been created and stored on a local computer where the organisation has installed the 
software and therefore only accessible on computers that have a trusted connection via VPN and secure 
credentials (certificates, RSA keys or complex passwords). This computer follows the security standards 
set by the ICS in compliance with current requirements.  
 
13. ACKNOWLEDGEMENTS  
The authors are grateful to the participants in this study. We also want to thank the Medical College of 
Barcelona, The Nurse College of Barcelona, Closca, Leroy Merlin, Barfim, Roche, for their personal 
protection equipment and health material donations. 
In addition, the researchers would like to thank Núria Prat Gil, Director of Primary Care at the 
Metropolitana Nord, and Josep Ma Bonet Simó, Assistant Director of Primary Care at the Metropolitana 
Nord.This project was funded by Departament de Salut. Generalitat de Catalunya . Call COVID19-PoC 
number SLT16_04 
 
14. COMPETING INTERESTS  
 
The authors declare no competing interests. 
 
15. LANGUAGE 
Case report forms must be completed in Catalan and Spanish and Inform Consent and Sheet participant 
information is available in English ,also.  
All written information and other material to be used by subjects and investigative staff must use 
vocabulary and language that are clearly understood. 
16.  COMPENSATION 
 
Participants will be covered by usually public insurance. They do not receive any compensation 
 

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17. OPEN ACCESS 
 
A public dissemination of the results will be made.  
 
18.PATIENT CONSENT DETAIL 
The informed consent and the patient information sheet are shown in appendix  
 
19. PROVENANCE AND PEER REVIEW NOT COMMISSIONED; EXTERNALLY PEER 
REVIEWED. 
Scientific Committee of The Foundation University Institute for Primary Health Care Research Jordi Gol i 
Gurina (IDIAPJGol) 
 
20. DATA SHARING STATEMENT 
The study data will not be openly available. If researchers or institutions are interested in the data, the 
authors will be able to release the data after asking the study participants for their consent and signing 
the relevant legal agreements allowing the data to be released to third parties. 
21. APENDICES 
The tables and figures cited in the text are shown below. 
 
 

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Table 1. ProHEpiC-19 Selection criteria. 
 
Inclusion 
criteria 
≥ 18 years of age 
Accept to take part in the 
study and sign the 
informed consent 
according to the 
Declaration of Helsinki. 
To be a health care 
professional  worker 
infected or exposed to 
SARS-CoV-2. 
Exclusion 
criteria 
< 18 years old 
Not to accept to take part 
in the study and/or not to 
sign the informed consent 
according to the 
Declaration of Helsinki. 
Not to be a health care 
professional worker 
exposed to SARS-CoV-2  
 
 

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Table 2 Participants assigned to each cohort 
Visit  Pre-cohorts enrolled 
Protocol 
0 
All possible participants 
Telephone screening/personal interview to explain the study and to decide if is an eligible 
participant and pre-cohort 
1 
Healthy exposed healthcare workers  
 
RT-PCR (SARS-CoV2)  
Anti-SARS-CoV-2 IgG and IgM antibodies (Nucleopcapside)   
Hemogram and serum biochemistry 
Recent infected healthcare workers  
 
RT-PCR (SARS-CoV2)  
Anti-SARS-CoV-2 IgG and IgM antibodies   
Hemogram and serum biochemistry 
T-cells and cytokines 
No-recent infected healthcare workers 
No visit 
2 
Recent infected healthcare workers  
Telephone symptoms control call 
Healthy exposed healthcare workers and 
No-recent infected healthcare workers 
 
No visit 
3 
Healthy exposed healthcare workers and 
No-recent infected healthcare workers 
 
No visit  
 
Recent infected healthcare workers with 
positive RT-PCR in Visit 1 or previously 
With positive RT-PCR in Visit 1 or previously: RT-PCR (SARS-CoV2) and Anti-SARS-CoV-2 IgG and IgM 
antibodies   
 
With negative RT-PCR in visit 1 and previously: Anti-SARS-CoV-2 IgG and IgM antibodies   
 
4 
Healthy exposed healthcare workers  
 
Anti-SARS-CoV-2 IgG and IgM antibodies   
 
Recent infected healthcare workers 
Anti-SARS-CoV-2 IgG and IgM antibodies   
T-cells and cytokines 
No-recent infected healthcare workers 
RT-PCR (SARS-CoV2)  

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Anti-SARS-CoV-2 IgG and IgM antibodies   
Hemogram and serum biochemistry 
 
5 
Healthy exposed healthcare workers  
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies   
Recent infected healthcare workers with 
positive RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
Recent infected healthcare workers with 
negative RT-PCR in visit 1 and previously 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies   
Non-recent infected healthcare workers 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies   
6 
Healthy exposed healthcare workers  
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies   
Recent infected healthcare workers with 
positive RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
Recent infected healthcare workers with 
negative RT-PCR in visit 1 and previously 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies   
Non-recent infected healthcare workers 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM antibodies   
7 
Healthy exposed healthcare workers  
Anti-SARS-CoV-2 IgG and IgM antibodies 
Recent infected healthcare workers with 
positive RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
 Recent infected healthcare workers with 
negative RT-PCR in visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
Non-recent infected healthcare workers 
Anti-SARS-CoV-2 IgG and IgM antibodies 
8 
Healthy exposed healthcare workers  
Anti-SARS-CoV-2 IgG and IgM antibodies 
Recent infected healthcare workers with 
Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 

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positive RT-PCR in Visit 1 or previously 
Recent infected healthcare with negative 
RT-PCR in visit 1 and previously 
Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
Non-recent infected healthcare workers 
Anti-SARS-CoV-2 IgG and IgM antibodies 
9 
Healthy exposed healthcare workers  
Anti-SARS-CoV-2 IgG and IgM antibodies 
Recent infected healthcare workers with 
positive RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
Recent infected healthcare with negative 
RT-PCR in visit 1 and previously 
Anti-SARS-CoV-2 IgG and IgM antibodies, T-cells and cytokines 
Non-recent infected healthcare workers 
Anti-SARS-CoV-2 IgG and IgM antibodies 
 

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Table 3 . Initial symptoms of COVID-19 
SARS-COV2 
CLINICAL 
SYMPTOMATOLOGY 
Headache 
Diarrhea  
Dyspnea 
Fever 
Cough 
Anosmia 
Arthralgias 
Asthenia 
Shivers 
Thoracic 
Epigastralgia  
Discomfort  
Myalgias  
Nausea  
Odynophagie  
Congestion 
 
 
 

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Table 4 Symptoms clinical monitoring*  
Symptom 
Date of 
onset of 
the 
symptom 
Duration 
(days) 
Observations 
Cardiovascular 
  
  
  
Fainting 
  
  
  
Pain/burning in chest 
  
  
  
Tachycardia 
  
  
  
Bradycardia 
  
  
  
Palpitations 
  
  
  
Visibly inflamed/ bulging veins 
  
  
  
Dermatologic 
  
  
  
COVID toe 
  
  
  
Dermatographia 
  
  
  
Other skin and allergy 
  
  
  
Peeling skin 
  
  
  
Petechiae 
  
  
  
Skin rashes 
  
  
  
Gastrointestinal 
  
  
  
Diarrhea 
  
  
  
Loss of appetite 
  
  
  
Vomiting 
  
  
  
Abdominal pain 
  
  
  
Nausea 
  
  
  
Constipation 
  
  
  
Gastroesophageal reflux 
  
  
  
Head/eyes/nose/ears/throat 
  
  
  
Runny nose 
  
  
  
Sore throat 
  
  
  
Hearing loss 
  
  
  
Other hearing/ear issues 
  
  
  
Tinnitus 
  
  
  
Vision symptoms 
  
  
  
Immunologic/autoimmune 
  
  
  
New allergies 
  
  
  
New anaphylaxis reaction 
  
  
  
Musculoskeletal 
  
  
  
Bone ache or burning 
  
  
  
Muscle aches 
  
  
  

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Tightness of chest 
  
  
  
Joint pain 
  
  
  
Muscle spasms 
  
  
  
Neuropsychiatric  
  
  
  
Acute confusion/disorientation 
  
  
  
Changes to sense of smell and taste 
  
  
  
Dizzines, unsteadiness or balance 
issues 
  
  
  
Hallucinations 
  
  
  
Headaches and related symptoms  
  
  
  
Insomnia 
  
  
  
Other sleeping symptoms 
  
  
  
Sleep apnea 
  
  
  
Slurring eord/speech 
  
  
  
All neurological sensations 
  
  
  
Brain fog 
  
  
  
Memory issues 
  
  
  
Neuralgia (nerve pain) 
  
  
  
Speech/language issues 
  
  
  
Tremors 
  
  
  
Vibrating sensations 
  
  
  
Pulmonary/Respiratory  
  
  
  
Dry cough 
  
  
  
Rattling of breath 
  
  
  
Breathing difficulty (normal O2 
saturation level) 
  
  
  
Cough with mucus production 
  
  
  
Coughing blood 
  
  
  
Other respiratory and sinus 
  
  
  
Shortness of breath 
  
  
  
Sneezing 
  
  
  
Reproductive/genitourinary/endocrine   
  
  
All menstrual issues 
  
  
  
Bladder control issues 
  
  
  

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Systemic 
  
  
  
Elevated temperature (98.8-100.4F) 
  
  
  
Fever (>100.4F) 
  
  
  
Chills/flushing/sweats  
  
  
  
Fatigue 
  
  
  
Low temperature 
  
  
  
Other temperature issues 
  
  
  
Post exertional malaise 
  
  
  
 
 
 
*Characterizing Long COVID in an International Cohort: 7 Months of Symptoms and Their Impact Hannah 
E. Davis1*, Gina S. Assaf1*, Lisa McCorkell1*, Hannah Wei1*, Ryan J. Low1,2*,Yochai Re’em1,3*,Signe 
Redfield1, Jared P. Austin4, Athena Akrami1,2*+ medRxiv preprint doi: 
https://doi.org/10.1101/2020.12.24.20248802 
 
 

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TABLE 5. Protocol to be followed according to the cohort of inclusion in the study 
 
Visit 
Study Cohorts 
Protocol 
0 
All possible participants 
Telephone screening/personal interview to explain the study 
and to decide if is an eligible participant and pre-cohort 
1 
 
Healthy Health Care Workers 
(HCW) 
 
RT-PCR (SARS-CoV2) 
Anti-SARS-CoV-2 IgG and IgM antibodies  
Hemogram and serum biochemistry 
Infected HCW 
           
RT-PCR (SARS-CoV2) 
Anti-SARS-CoV-2 IgG and IgM antibodies  
Hemogram and serum biochemistry 
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
Healthy HCW 
No visit 
2 
Infected HCW       
Telephone symptoms control call 
3 
Healthy HCW 
  
No visit 
  
Infected HCW      with positive 
RT-PCR in Visit 1 or previously 
With positive RT-PCR in Visit 1 or previously: RT-PCR (SARS-
CoV2) and Anti-SARS-CoV-2 IgG and IgM antibodies  
  

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with negative RT-PCR in visit 1 and previously: Anti-SARS-CoV-2 
IgG and IgM antibodies  
  
 Healthy HCW 
No visit 
  
4 
Healthy HCW 
  
Anti-SARS-CoV-2 IgG and IgM antibodies  
  
Infected HCW      
Anti-SARS-CoV-2 IgG and IgM antibodies  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
Healthy HCW 
RT-PCR (SARS-CoV2) 
Anti-SARS-CoV-2 IgG and IgM antibodies  
Hemogram and serum biochemistry 
  
5 
Healthy HCW 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies  
Infected HCW      with positive 
RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 

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 Infected HCW      with negative 
RT-PCR in visit 1 and previously 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies  
Healthy HCW 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies  
6 
Healthy HCW 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies  
Infected HCW      with positive 
RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
Infected HCW      with negative 
RT-PCR in visit 1 and previously 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 

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without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies  
Healthy HCW 
with antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
without antibodies in Visit 4: Anti-SARS-CoV-2 IgG and IgM 
antibodies  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
7 
Healthy HCW 
Anti-SARS-CoV-2 IgG and IgM antibodies 
Infected HCW      with positive 
RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
 Infected HCW      with negative 
RT-PCR in visit 1 and previously 
Anti-SARS-CoV-2 IgG and IgM antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
Healthy HCW 
Anti-SARS-CoV-2 IgG and IgM antibodies 
8 
Healthy HCW 
Anti-SARS-CoV-2 IgG and IgM antibodies 
Infected HCW      with positive 
RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 

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Infected HCW      with negative 
RT-PCR in visit 1 and previously 
Anti-SARS-CoV-2 IgG and IgM antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
Healthy HCW 
Anti-SARS-CoV-2 IgG and IgM antibodies 
9 
Healthy HCW 
Anti-SARS-CoV-2 IgG and IgM antibodies 
Infected HCW      with positive 
RT-PCR in Visit 1 or previously 
Anti-SARS-CoV-2 IgG and IgM antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
Infected HCW      with negative 
RT-PCR in visit 1 and previously 
Anti-SARS-CoV-2 IgG and IgM antibodies,  
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 
 
Healthy HCW 
Anti-SARS-CoV-2 IgG and IgM antibodies 
T-cells  (INFg Elispot assays, Combinatorial flow-cytometry) 
Citokines multiplex assay 

ProHEPIC-19 Study Protocol V1.1                                                                                 4R20-105 
 
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Figure 1.Decision criteria for assigning participants to cohorts 
DOES THE PARTICIPANT IS/WAS INFECTED BY SARS-CoV-2 
YES 
NO 
HEALTHY HEALTH 
CARE WORKERS 
COHORT
≥15 DAYS FROM RT-PCR OR COMPATIBLE 
COVID-19 SYMPTOMS? 
YES 
NO 
NON-RECENT HEALTH 
CARE WORKERS 
COHORT
RECENT HEALTH 
CARE WORKERS 
COHORT

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INFORMED CONSENT 
 
Study title: SARS-CoV-2  infection among healthcare professionals: demographic 
characteristics and serological and immune responses related to progression's 
phenotype. 
 
I, ............................................................................................................. 
I have read the foregoing information sheet to the patient  and / or participant. 
I have had the opportunity to ask questions about  the study. 
I have received enough information about the study. 
I have talked to:  
I understand that my participation in this study is completely voluntary. 
I understand that I can withdraw from the study: 
1. When I want it, 
2. Without giving explanations, and 
3. Without affecting the health care that I will receive. 
In accordance with the provisions of Regulation (EU) 2016/679 of the European Parliament 
and the Council of April 27 on Data Protection (RGPD) and Organic Law 3/2018, of 
December 5, on data protection and guarantee of digital rights, I declare to have been 
informed of my rights, of the purpose of collecting my data and of the recipients of the 
information. 
I consent voluntarily to participate as a participant in this research. Date (day/month/year) 
……………………………………….. 
Name of Participant (Name and surname) 
 ………………………………… 
Signature of Participant 
 ……………………………………… 
 
 
Name of the clinical researcher  
………………………………………… 
Signature of the clinical researcher 
………………………………………… 

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PATIENT/PARTICIPANT INFORMATION SHEET  
Introduction 
This Information Sheet is for people who are invited to participate in the research entitled: “SARS-
CoV-2 infection among healthcare professionals: demographic characteristics and serological 
and immune responses related to progression's phenotype.”.  
Name of principal investigator: Pere Torán Monserrat / Concepción Violan Fors  
Name of the organisation: IGTP / IDIAPJGol  
You will receive information about this research and will be invited to be part of it. Before you 
decide, you are welcome to talk to anyone you feel comfortable with about the research. If there is 
anything you don't understand, don't hesitate to ask and we will review the information.  
Research objectives  
At the moment, healthcare professionals are a group at high risk of contracting SARS-CoV-2 
infection. The evolution of the SARS-CoV-2 pandemic suggests that epidemic waves may recur 
within a short period of time since the factors that condition the evolution of the infection, the 
permanence of the virus and the immunity of infected individuals are largely unknown. In addition, 
healthy carriers are not clearly identified. The main objective of this research is to consolidate a cohort 
of health professionals exposed to SARS-CoV-2, to measure the serological and immunological 
characteristics that allow distinguishing symptomatic from asymptomatic carriers and to compare it 
with a cohort of patients treated in the same health professionals' workplaces. 
Type of research and procedures  
This is a prospective study with three cohorts, two of them consisting of healthcare workers and one 
consisting of patients. This research will involve the collection of biological samples for subsequent 
analysis, as well as the collection of socio-demographic and quality of life data:  
1. At least two swab samples shall be taken for the RT_PCR diagnostic test of oropharyngeal and 
nasopharyngeal exudates to detect the presence of the virus. The first sample shall be taken when 
included in the cohort, the second sample 14 days after inclusion in the cohort and it may be 
necessary to obtain some more samples until the absence of virus is verified. 
2. Two capillary blood samples obtained by finger prick will be taken on the same days as the exudate 
test for RT_PCR. This test is performed to measure C-reactive protein in blood as a marker of the 
degree of inflammation present in the organism.  
3. Six blood samples will be taken for serological and immunological tests obtained by venous 
puncture. The first one at inclusion in the cohort, the second on day 14 after the first RT-PCR, the 
third at day 60, the fourth 90 days after the first RT-PCR test, the fifth between days 180 and 270 

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and the sixth at the end of the study, i.e. one year after the inclusion visit. 
4. A questionnaire on demographic, clinical, social and psychological data will be administered during 
the first month after inclusion in the study. Additionally, some of the participants will be 
contacted afterwards with the sole purpose of offering them the opportunity to complement the 
information on the psychosocial impact of SARS-CoV-2 by means of an individual interview. 
This would only be carried out after obtaining a new ad-hoc informed consent, designed and 
validated by the reference Research Ethics Committee (REC).   
Selection of participants  
We are inviting 600 healthcare workers included in the protocol for assistance to people in isolation 
due to SARS-CoV-2 exposure, plus 300 additional health professionals with different risks of 
infection, systematically selected from the Northern Metropolitan Territorial Management of the 
Catalan Health Institute (ICS).  
Voluntary participation  
Your participation in this research is completely voluntary. It is entirely your choice whether or not to 
participate.  
Benefits  
The results of this study will contribute to the characterisation of an algorithm to identify carrier status 
and immune response in healthcare professionals with various levels of exposure to SARS-CoV-2. 
This will optimise the management of human resources with sound criteria for the safety of 
professionals and patients, thus strengthening the health system to cope with this pandemic and the 
new waves predicted. You will have the results of the tests and will be able to know your degree of 
immunisation against the virus. 
Processing of biological samples  
First of all, you should know that donating your samples is voluntary. You are under no obligation to 
donate your samples in order to receive medical care. Even if you agree now, if you change your mind 
later, you can request that any samples that have not yet been analysed are destroyed.   
Purpose of sample collection  
The samples you donate will be used for current or future studies related to COVID-19 infection. 
Your samples may only be used for the purpose stated in this information sheet. In the event that they 
may be used for other non-COVID-19 related research in the future, they must be approved by an 
Ethics Committee and you will be asked for a new consent form.  
 
 

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Where will the tests be carried out?  
Your samples may be analysed in different laboratories of this institution, although they may be 
analysed in other centres, or even in foreign laboratories, depending on the types of analysis to be 
carried out.   
What will happen to my samples once they have been analysed?  
Your samples may only be used for the purpose stated in this information sheet. Your samples may be 
used in ongoing studies of COVID-19 infection, but may also be retained for future research related to 
COVID-19 infection.   
All studies using your samples must be approved by a Research Ethics Committee. You may request 
to be informed about the studies in which your samples have been used.   
Your samples will be stored mainly in the Germans Trias i Pujol Health Sciences Research Institute 
(IGTP) Biobank, an institution that has been accredited by the health authorities specifically to store 
samples for research under the conditions required by law. The treatment and use of all samples will 
be carried out in accordance with the Biomedical Research Act (14/2007) and Royal Decree 
1716/2011, which regulates biobanks. The samples will be kept for 15 years. They may be transferred 
to third parties if the current Biobank legislation is complied with and they will not be sold under any 
circumstances.  
 
In some cases, they may become part of sample collections accredited by the health authorities. In this 
case, they can neither be transferred to third parties nor sold, but they could later be integrated into the 
Biobank of our centre.  
The samples will be identified in a coded form. This means that the information on the labelling of the 
samples cannot be linked to the person who donated the sample. Please see below the section on 
Confidentiality and data protection.  
Confidentiality  
The information we collect in this research project will be confidential and no one other than the 
researchers will be able to see it. Once all the information has been obtained, it will be transferred to a 
database in which an encrypted identification code will be created and no information that would 
allow your identity to be recognised will be recorded.  
It is also important for you to know that this proposal has been reviewed and approved by the 
Research Ethics Committee of the IDIAP Jordi Gol and the Germans Trias i Pujol Research Institute 
(IGTP).  

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The processing of your personal data will comply with the provisions of the Organic Law 3/2018, of 5 
September, on the Protection of personal data and guarantee of digital rights and Regulation (EU) 
2016/679 on the protection of natural persons that relate to the processing of personal data and the 
free movement of such data. In accordance with the provisions of the aforementioned legislation, you 
may exercise the right of access, rectification, erasure, restriction of processing, portability and 
opposition to processing.  
Who to contact  
If you have any questions, you can ask them now or later, even after the study has started. In any case, 
you can contact the project managers: Pere Torán Monserrat ptoran.bnm.ics@gencat.cat, Concepció 
Violan Fors.  cviolan@igtp.cat  
If you agree to participate in this study, please express your consent by completing the document 
available below.