Screening for Hepatitis c in People Who Inject Drugs in Armenia-Colombia
Conditions: Hepatitis C Virus Infection
Sponsor: Asociación Colombiana de Hepatología

 
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SCREENING FOR HEPATITIS C IN PEOPLE WHO INJECT DRUGS IN  
ARMENIA-COLOMBIA 
 
A. TECHNICAL APPROACH 
Problem statement and justification 
Hepatitis C virus (HCV) infection causes acute and chronic liver disease and can lead to 
cirrhosis, liver failure, or hepatocellular carcinoma. Globally, an estimated 58 million people 
are chronically infected with hepatitis C virus, with around 1.5 million new infections per year 
(1). The World Health Organization (WHO) estimated that in 2019, approximately 290,000 
people died from hepatitis C, mainly from cirrhosis and hepatocellular carcinoma (primary 
liver cancer) (1). 
Despite undeniable advances in the treatment of hepatitis C infection and the WHO strategy 
to eliminate hepatitis C by 2030 (2), this infection continues to be a major public health 
problem globally, for which there is no vaccine, and access to diagnosis and treatment is 
low, however, antivirals can cure more than 95% of cases of this infection (1). Acute HCV 
infection is usually asymptomatic, and it is estimated that between 15% and 45% of infected 
people clear the virus spontaneously without any treatment. The other cases (55% to 85% 
of people) develop chronic HCV infection, and among these, there is a high (15% to 30%) 
estimated risk of liver cirrhosis in the absence of therapeutic intervention (1). Due to the 
largely asymptomatic nature of HCV infection, many HCV-positive individuals are unaware 
of their HIV status. Others have been diagnosed but cannot get treatment. Lack of rapid and 
reliable diagnosis and high cost and limited access to treatment are barriers to disease 
control and elimination (3). 
People who inject drugs (PWID) are at increased risk for HCV. Several studies have reported 
high HCV prevalence rates, especially among PWID (4–6). Worldwide, there are around 11 
million PWIDs (7). Approximately 39.4% have viremic HCV infection (8). It is estimated that 
between 23% and 39% of new HCV infections occur among PWID, and one in three HCV 
deaths are attributable to injection drug use (9). In some regions, such as Eastern Europe 
and Central Asia, HCV prevalence rates are particularly high. Additionally, there are 
approximately 2.3 million coinfections between HIV and HCV worldwide, of which more than 
half (1.3 million) occur in PWID (7). The coexistence of these two health conditions leads to 

 
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accelerate the progression of liver disease and complicate treatment for HIV. The global 
prevalence of HCV in 2019 among PWID was 50.2%, which is equivalent to 5.6 million 
people who inject drugs and live with hepatitis C. Four countries (Brazil, China, Russia, and 
the United States) have most people with recent injection drug use living with HCV. 
Together, these countries represent more than half (51%) of all people with recent injection 
drug use living with HCV worldwide (10). 
PWID are usually exposed to a higher risk of various infectious diseases, mainly due to their 
drug consumption behaviors and habits, in addition to the risks and harms associated with 
the respective routes of self-administration (11,12). Needle sharing among intravenous drug 
users is a major risk factor for HCV infection (13). The behavior of illicit drug users may 
increase the risk of viral infections through choice of drug, frequency of use, and sharing of 
paraphernalia, as well as unprotected sexual activity, multiple sexual partners, and 
prostitution (25). Several studies indicate that drug users have played a key role in the global 
spread of HCV (5.14). 
In Colombia, the prevalence of hepatitis C among PWID has been measured locally in some 
cities of the country. The first measurement was carried out with the 2002 study in Bogotá. 
Later, in 2014 it was held in Bogotá, Pereira, Medellín, Cucuta and Armenia. The figures at 
that time already indicated a clear impact by this virus in this population. In Bogotá it went 
from 1.7% in 2002 to 6.7% in 2014. The cities with the lowest prevalence in 2014 were 
Cucuta with 21.4% and Armenia with 30.9%, while in Medellín the estimated prevalence It 
was 35.6% and in Pereira it was 44.4% (15). For 2021, the prevalence of hepatitis C was 
measured in Bogotá, Medellín, Santiago de Cali, the metropolitan area of Pereira-
Dosquebradas, Medellín, Cucuta and Armenia (15,16). The results of prevalence of 
antibodies against hepatitis C were as follows: Cali with 80.2%, is the city with the highest 
reactivity, followed by Pereira and Dosquebradas with 71.4%, Armenia with 69.6% and 
Cucuta with 62.8% (15,16). In Medellín, antibody reactivity was found in 32.7% of PWIDs 
and in Bogotá in 10.7% of them. The comparison of the findings between this study and the 
CES University study in 2014 shows a slight reduction in the hepatitis C figures in Medellín, 
and an increase of several percentage points in Bogotá, Pereira –Dosquebradas, Cucuta 
and Armenia. There is no previous data on hepatitis C for Santiago de Cali. 
With the promising effectiveness of antivirals to treat the infection and prevent transmission 
(17), WHO declared the intention to eliminate viral hepatitis by 2030 (18). Targets for 

 
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elimination include an 80% decrease in new HCV cases, 90% of infections diagnosed, 80% 
of infections receiving treatment, and 65% reduction in HCV-related deaths. However, most 
people with HCV are unaware of their status due to asymptomatic infection, low screening, 
and, among those screened, limited diagnosis and linkage to care (19,20). The lack of 
accurate national HCV surveillance systems, particularly for key populations such as PWID, 
contributes to the challenges of HCV control and elimination (21). Additional studies are 
needed to improve HCV screening, evaluation, and treatment to reduce the burden of HCV 
infection among PWID. 
In accordance with the above, the objective of this study is to estimate the prevalence of 
anti-HCV antibodies in PWID between 18 and 65 years of age in the municipality of Armenia 
(Quindío) through public health strategies such as screening that allows characterize the 
affected people and facilitate access to diagnosis and treatment. 
Theoretical framework and state of the art 
Hepatitis C has constituted a major public health problem worldwide, causing liver failure, 
cirrhosis, hepatocellular carcinoma, and acute and chronic hepatitis C infections with a high 
mortality rate. Although risk factors commonly associated with the transmission of HCV 
infection include blood transfusion from unscreened donors, unsafe therapeutic injections, 
and other healthcare-related procedures, the majority of new and existing infections in most 
countries have occurred as a result of injection drug use (22). 
PWID engage in high-risk behaviors and are at risk of contracting or transmitting viral 
infections such as human immunodeficiency virus (HIV) and hepatitis. That's because 
viruses are transmitted through blood and other body fluids. Generally, contagion occurs in 
two ways: a) when a person injects drugs and shares needles, or other items used for 
consumption and b) when drugs affect judgment, and the person has unprotected sexual 
relations with a partner. infected. This can happen to men and women alike (23,24). 
Among PWIDs, the main route of transmission is through the exchange of drug preparation 
and injection equipment (for example, syringes, needles, filters, etc.) (25). HCV is resistant 
and is able to survive on drug preparation equipment (e.g. needles, syringes, filters, etc.) for 
several days or weeks (26). Furthermore, the risk of HCV transmission is greater than (1) 
HIV infection, consistent with greater transmission from exposure to contaminated injections 

 
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(2.5-5.0% for HCV vs. 0.5%-2.0% for HIV), and a higher prevalence of HCV than HIV among 
people who inject drugs (and therefore risk of exposure) (27). 
WHO has identified strategies for the prevention, detection and treatment of infections 
caused by HCV and other hepatitis viruses worldwide to reduce the number of such 
infections (1). Better detection of undiagnosed cases, greater availability of treatment and 
better coordination of care are important components of the elimination strategy (1). In 
addition to primary prevention strategies such as opioid replacement therapies and needle 
and syringe programs, early diagnosis, and treatment of HCV in PWID, which is considered 
a high-risk group, will help reduce complications and mortality. HCV-related mortality, 
especially in PWID. The “treatment as prevention” approach is gaining importance in 
treatment guidelines, because eradication of HCV infection through early diagnosis and 
treatment would impact both individual well-being and reduce the spread of HCV infection 
the disease in PWID (28–30). PWID have been considered a difficult group to reach, 
manage, and treat because HCV treatment management in these individuals is challenging 
and they have a higher risk of reinfection (31). 
Initially, HCV treatment guidelines excluded PWIDs from consideration, citing concerns 
about adherence, increased susceptibility to side effects, and reinfection (32). However, 
there is now compelling evidence that HCV treatment is safe and effective among PWID 
(33). International guidelines now recommend treatment for PWID after individualized 
evaluation (3. 4). 
Any attempt to avert the threat to public health care posed by the impending burden of HCV 
among PWID will urgently require innovative changes to alter the currently inefficient system 
for care of HCV infection among this vulnerable population. A relevant expansion of 
treatment among PWID is impossible without massively reducing barriers to care. Low 
awareness, among patients, healthcare providers, policy makers, political leadership and 
the general public, as well as discrimination and stigmatization of PWID are the main barriers 
for people who inject to access the HCV care (35,36). Many of these barriers are the result 
of the criminalization of drug use (37). Repressive drug policy is hindering effective public 
health measures for people who inject drugs and, therefore, increasing the HCV and HIV 
epidemic in this population (38). 

 
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Objectives  
General objective 
To estimate the prevalence of hepatitis C antibodies through rapid test screening and 
identify the behaviors associated with infection in people between 18 and 65 years of age 
who inject drugs in the city of Armenia. 
Specific objectives 
• 
Estimate the size of the population of people who inject drugs in the city of Armenia. 
• 
Estimate the prevalence of antibodies against HCV in PWID. 
• 
Socio-demographically characterize the study population. 
• 
Describe the epidemiological characteristics of PWID with positive antibodies for HCV. 
• 
Identify behaviors associated with HCV infection among PWID. 
• 
Determine the proportion of PWID with positive HCV antibodies and positive HCV viral 
load. 
• 
To carry out an exploratory analysis of the association of positive cases and the different 
risk factors for HCV infection among PWID. 
• 
To quantify how many PWID with positive viral load for HCV were able to access the 
management route and treatment. 
B. METHODOLOGICAL APPROACH 
Type of study 
An anonymous, confidential analytical cross-sectional study will be carried out, in which the 
prevalence of hepatitis C will be estimated, the behaviors associated with the consumption 
of psychoactive substances by injection will be analyzed in people between 18 and 65 years 
old who inject drugs in the city of Armenia. 
 
 

 
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Population and sample 
Population 
The universe for this study was people between 18 and 65 years old who have injected 
drugs in the last six months in Armenia and the participants were selected through the 
implementation of the respondent-driven sampling (RDS). 
Inclusion and exclusion criteria 
Inclusion criteria 
• 
Be between 18 and 65 years old. 
• 
Having injected psychoactive substances in the last six months. 
• 
Residing in the city or metropolitan area of study in the last six months 
• 
Present the invitation coupon. 
• 
Person with Colombian nationality, Venezuelan migrant population. 
Exclusion criteria 
• 
Person who, due to physical, cognitive or limitations derived from the use of 
psychoactive substances, is not able to answer the survey autonomously. 
• 
Person who is in a state of consumption and is prevented from responding to the survey. 
• 
Person who refuses to collaborate in the study. 
Sampling frame 
Statistical units 
The statistical units of analysis, observation and sampling are the key population of Armenia. 
Target population size 
Sample sizes and precision for estimating a population proportion. 
Population size (for finite population correction factor or fcp) (N): 5801 
Hypothetical % frequency of the outcome factor in the population (p): 77.80%2+/-5 
Confidence limits as % of 100(absolute +/-%) (d): 5% 
 
1Patients treated at the Armenia City Listening Center between 2016-2023. 
2Estimated based on the 2021 prevalence (62.80%), along with the growth speed of 5% per year. 
 

 
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Design effect (for group surveys-EDFF): 1 
Sample size (n) for various confidence levels. 
Interval 
Trust (%) 
Size 
of the sample 
95% 
186 
80% 
98 
90% 
145 
97% 
213 
99% 
260 
99.9% 
331 
99.99% 
377 
Sample size n = [EDFF*Np(1-p)]/ [(d2/Z21-α/2*(N-1) + p*(1-p)] 
 
Source: Results from OpenEpi, version 3, the SSPropor open-source calculator 
The sample size may vary by up to 10%, taking into account (i) the methodological 
adjustments that are made once the study begins, in which case the sample size will be 
adjusted. 
Sample size 205 individuals. 
Sample design 
Due to the type of population (PWID), it is not possible to have a known sampling frame 
that allows designing a conventional probabilistic sample, so a chain reference sample will 
be taken based on those who respond (respondent driven sampling -RDS), a method that 
has proven its usefulness to access hard-to-reach populations (15,16). Respondent-driven 
sampling is a network-based sampling method, which makes it possible to draw statistically 
valid samples from previously unreachable groups. It is based on long referral chains and 
therefore several waves of recruitment. Likewise, several waves of recruitment make it 
possible to ensure that the final sample is independent of the seeds and this sample is 
similar to the population of the network from which the sampling was done (16). For 
example, in the prevalence study in 2021, the method estimated a population of 463 (95% 
CI: 431-497) people who inject drugs. Regarding completeness, through the RDS 
methodology, 53.4% of the population were identified and the method estimates that 
80.19% of the population in the city of Armenia was identified3 (16). 
The seeds are those leaders of the population who begin the information gathering process. 
 
3https://www.minjusticia.gov.co/programas-co/ODC/Documents/Publicaciones/Informe%20de%20Resultados%20de%20Investigacio%CC%81n%20PID%20Armenia-
Cu%CC%81cuta%20Marzo%202022. pdf 

 
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In this sampling it is necessary to link the referral with the interviewee, which is achieved 
through coupons marked with the RDS code (unique number) that allows identifying all the 
referrals of a seed and therefore its social network. Likewise, the reference quota is 
controlled with a maximum number of coupons, which for this study would be three per 
interviewee, which allows limiting “recruiter” bias; coupons are an inclusion criterion that 
allows determining whether the person was eligible for participate and essential to preserve 
the link between referent and referred. 
Respondents recruit their peers, as in network-based samples, and researchers keep track 
of who recruited whom and their number of social contacts. A mathematical model of the 
recruitment process then weights the sample to compensate for non-random recruitment 
patterns. This model is based on a synthesis and extension of two areas of mathematics, 
first-order Markov chain theory (i.e., sampling occurs with replacement, the sample 
becomes independent of the seeds). And biased network theory, which were not part of 
the standard toolset of mathematical sampling theory. The resulting statistical theory, called 
RDS, allows researchers to provide unbiased population estimates and measures of the 
precision of those estimates. 
Variables 
The dependent variable would be the identification of HCV antibodies, which is qualitative 
with nominal measurement as presence (positive) or absence of the virus (negative). The 
questions that will make up the survey will be the independent variables, whose nature is 
qualitative or quantitative and the measurement levels are nominal and ratio, such as age, 
number of times per day in which you inject, number of people with whom you have shared 
syringes and needles, injection equipment, number of casual or commercial couples, among 
others. 
Collection of information and sampling procedure 
Once approval has been obtained from the research ethics committee for this study, in the 
first phase people called seeds are selected, a record of the identified population will also 
be made, and the first part of the capture-recapture methodology will be applied. In the 
second phase of expansion, it is expected to recapture the population and estimate the 
population, with which the waves of the RDS methodology are carried out. 
The researcher initially searches for individuals (called seeds) who belong to the population 

 
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of interest and who know a measurable number of people in that population. The seed is 
contacted, invited to the headquarters, the survey and the defined biological samples are 
applied, and they are given the three coded coupons that also contain the information about 
the study, the headquarters, and the opening hours. Each seed will deliver the coupons to 
known people who meet the eligibility criteria. Once the seeds have delivered their coupons 
to their peers, they are expected to come to the headquarters to participate, once the 
information collection process is completed. In this way, the enrollment process is 
developed in waves until the predefined sample size is reached. Each coupon contains 
information about the headquarters, including the address, opening hours and contact 
telephone number, as well as the RDS code assigned to each participant, which allows 
identifying who recruited whom and reconstructing the chain of referrals or recruitment. 
Potential participants will be presented with the purposes of the research, and it will be 
asked if they are interested in participating. The informed consent will be read and 
explained to them, and they will subsequently be asked to sign the consent. The 
standardized information collection phase will then proceed. These individuals will undergo 
pre-test counseling, a presumptive rapid test for HCV, and if positive, contact information 
will be requested for post-test counseling. 
The collection of information on social and demographic variables, risk factors associated 
with HCV infection, and access to the health system will be carried out with the application 
of a structured survey conducted by a previously trained interviewer. You will explore social 
and demographic characteristics, social networks, drug use and injection, sexual behavior, 
sexually transmitted infections, HCV knowledge, and rapid test results and viral load. 
The sample will be taken by trained personnel (indications for storage, application and 
reading of each test), through capillary puncture of the pad region of the fingers of the right 
hand, after asepsis and antisepsis, the rapid HCV diagnostic test will be used. Ab Plus 
Rapid test with INVIMA registration 2018 RD-0002353-R1 with operational characteristics 
that allow it to be performed at the point of care, obtaining a result in 15 to 20 minutes. This 
rapid HCV antibody test may be replaced by another with similar operating characteristics 
(sensitivity and specificity). The recommendations described by the test manufacturer will 
be followed (Annex 1). If it is reactive, it will be defined as a presumptive result because it 
is a screening test that requires confirmation of diagnosis, through a ribonucleic acid (RNA) 
detection test (viral load) in accordance with the provisions of the Hepatitis C Guide of the 

 
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Ministry of Protection Social (2019) to establish the diagnosis of chronic infection and in 
the case of obtaining a positive result in the latter, the HCV infection will be confirmed, and 
the participants will be channeled to health services for the respective treatment through of 
your EAPB. 
All information obtained will be recorded and processed individually (case by case) through 
a digital information system with backup in the cloud in real time using a Case Report Form 
(CRF) capture format, with the use of a tablet. This will allow daily monitoring of the 
behavior of RDS sampling. The information will be guarded and encrypted for the exclusive 
use of the main researchers belonging to the Colombian Association of Hepatology, a final 
report will be presented. The information will be anonymized for your use. 
Quality assurance procedure 
To control information biases, an instrument applied and reviewed in previous research, and 
which is part of the updated protocols for these studies will be used; Likewise, as a measure 
to control information bias in interviewers, training will be carried out in the proper handling 
of the collection instrument. In addition to the above, the coordinators will review the 
interviews collected by the interviewers to corroborate their correct completion. In the 
process of controlling the interviewee's information biases, the confidentiality of the 
information provided by the population is guaranteed. 
To control selection bias, the recognized sampling methodology of “Respondent Driven 
Sampling” is used. With the selection of the seeds, special care is taken to identify the 
properties of the social network (identification of subgroups, connectivity, personal network); 
the acceptability of the study (interest, possible reasons for not participating); the 
identification of seeds (large and diverse social network, leaders or well-known, support the 
study, enthusiastic); and take special care of logistics (site, schedules, coupon design, staff 
profile). 
Type of error/bias 
Description 
Control 
Selection bias 
Include 
people 
who 
meet inclusion criteria. 
A short screening-type questionnaire will be 
designed to identify people. 
The person performing the screening must be 
experienced. 
Compliance 
with 
study 
inclusion/exclusion 
criteria. 

 
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Precise definition of the objectives and scope of 
the study to field personnel. 
Pilot test. 
Information bias 
Data quality 
Source of information 
Intra and interobserver 
errors 
Unanswered questions 
 
 
Staff training. 
Application and evaluation of the pilot test. 
Supervision of the personnel who carry out the 
collection. 
Personnel with experience on the subject. 
Pilot test 
To implement the research protocol, ten people will be included (1 seed and 9 referrals), 
who inject drugs, the collection instrument will be applied where the clarity of the instrument 
will be evaluated and the field work manual will be adjusted (terms used, coherence of the 
content, order of the questions), the time of application, the disposition of the interviewers, 
the ability to generate confidence in the participants and finally the acceptance of the 
respondents. After carrying out the pilot test, it is expected to obtain the final version of the 
instrument and procedures. 
Data analysis plan 
The RDS itself estimates the sampling errors described through measures such as 
homophily, heterophily, and proportional estimates of the population. Initially, homophily will 
be calculated, which describes the extent of the ties of the social network inside or outside 
it. The homophily/heterophily scale is between 1 and -1, where 1 indicates that all 
participants with certain characteristics invited someone with the same characteristics, while 
-1 indicates that all participants with a certain characteristic invite someone with one 
characteristic. opposite or different; 0 (zero) was the point at which participants randomly 
invited other individuals from the population of all possible invitees. 
The RDS technique uses the information available in the sample about the social network 
to obtain asymptotically unbiased estimators of the proportions of the population in the 
different groups in which they want to be characterized. These estimators are called 
population prevalence estimators. 

 
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Descriptive statistics (frequencies and percentages) will be estimated. For continuous 
quantitative variables, measures of central tendency (average and median) and measures 
of dispersion (variance, standard deviation) will be estimated. 
A descriptive analysis of the characteristics of the population under study will be carried out, 
according to the socio-demographic variables: sex, age, educational level, stratum, 
insurance, in addition the prevalence of HCV and the proportions of the sample and the 
population will be obtained. according to the risk behavior variables associated with 
infections. All these analyzes are carried out by estimating the population with their 
corresponding 95% confidence intervals. 
Possible relationships between sociodemographic variables, risk factors and documentation 
of positive antibodies for HCV will be explored. For this purpose, a bivariate analysis will be 
proposed using chi-square tests to explore the proposed associations. The test allows us to 
determine, for each pair of variables, the existence or not of the relationship through the 
difference between the value that should result if the two variables were totally independent, 
and the one observed in the data. The greater the difference, the greater the relationship 
between the variables. 
The significance between the relationships will be determined by the p-value of the test, a 
value less than 0.05, it is accepted that there is a statistical association between the two 
variables analyzed. 
The processing will be carried out in the RDSAT® version 5.6 program. 
C. WORK PLAN 
Phase 1. Exploratory component 
With the purpose of making an approach and identifying barriers to the study and possible 
alternative solutions to consumption contexts, as well as establishing inter-institutional 
alliances that facilitate communication with research subjects called seeds, an exploratory 
phase will be carried out through the use of a qualitative technique that enables the collection 
and construction of data that enriches the information obtained by the subsequent 
application of the structured survey that underpins the present study. 
We will seek to identify characteristics of the social network: subgroups, connectivity, 
personal network, the acceptability of the study, interest, possible reasons for not 

 
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participating, as well as define logistical aspects, such as the site for collecting information, 
schedules, coupon design and the profile of the personnel who will serve the prioritized 
populations. 
Through interviews with representatives of various institutions in Armenia who, from 
research, care, and treatment, offer the research team a general overview of the context 
and care routes; and from there provide elements that guide field work and the analysis of 
the information obtained from the surveys. 
After prioritizing institutions, they will be inquired about the description of the problem, 
characterization of the populations, institutional experiences, research, territorial location, 
methods of approach, risk behaviors and practices for HCV infection, among others. The 
estimated time to complete the exploratory phase will be two months, this includes the 
collection, analysis and interpretation of the data and the understanding of the conditions 
found. 
Interviews with key informants 
These people are considered experts in the social and cultural context of PWIDs, with the 
objective of understanding the context and behavior of users; example of key informants: 
people who have worked with populations, people from institutions, NGOs, among others. 
In this first phase, the capture technique, already described previously, will also be applied. 
Phase 2. Quantitative component 
Hidden populations have two basic characteristics: first, they lack a sampling frame, so their 
size and real margins are unknown; and second, the people who belong to them are 
especially wary of offering information to researchers since they normally follow stigmatized, 
frowned upon or illegalized behaviors, therefore, locating them can be difficult. 
This limitation is intended to be resolved with the RDS sample design, by applying a coupon 
system that limits the possibility for each informant to select no more than three informants, 
to nullify the biases caused by the presence of semi-professional recruiters or by a desire 
for super-collaboration of some people that causes an overrepresentation of the networks 
of a specific individual. Social support vouchers will be used for respondents to promote 
inclusion and diversity, which encourage them to have other referrals and they must travel 
to a place to carry out the entire collection and sampling procedure. 

 
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The seed must be very well selected, with whom respect for privacy, confidentiality, and 
respect for autonomy will prevail, explaining that the coupon system will become effective 
when the referred person agrees to go to the predetermined place to carry out the collection 
procedure. of information and the social support bonus system, aims to recognize the costs 
generated by transportation, transfer, informing their social networks about the 
characteristics of the project and that this could in no way affect them due to the nature of 
confidentiality. of this research. 
Field work (quantitative phase) 
This stage begins with the process of selecting the interviewers to collect information and 
who meet the requirements of education, training and work experience; It is expected to 
have a nursing assistant for the collection process; and will be trained in voluntary testing 
for HCV; It is proposed that interviewers can carry out different activities such as screening, 
identification of injecting drug users (IDU) and reception tasks. At the same time, the 
instrument is reviewed and adjusted. 
Once the first version of the adjusted survey is approved, the pilot test will be carried out, to 
test the methodological design, calculate the completion time, adjust the project schedule, 
verify the required attitude of the staff and the most appropriate techniques. for quality 
control. 
The work team will have the following elements to carry out the field work: identification 
badge, institutional information, informed consent, survey, and pen, in addition to the 
implements for taking blood samples. 
Instructions for the personnel who carry out the collection. 
The personnel who carry out field work have five days of prior training; In the training, the 
procedures will be reviewed, and exercises will be carried out seeking to practice what has 
been learned. This training should be used to ask questions about any procedure to avoid 
errors in actual counseling and interviews. 
During the collection process, we will insist on the importance of having absolute knowledge 
of the topic, conducting the surveys in private, asking questions exactly as the question is 
formulated in the survey. Asking the questions slowly to make sure the person understood 
(in people who require it). 

 
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According to previous studies, completing the survey will not last more than approximately 
60 minutes. The entire information collection process that includes informed consent, 
application of the survey, pre-test counseling and sample collection will last approximately 
one hour and thirty minutes. 
Throughout the process, one of the researchers with extensive experience in the topic of 
public health surveillance will coordinate the actions on the ground with personnel, also with 
experience in the collection process; That is to say, during the training-concertation-study 
process of HCV seroprevalence -post-test counseling, monitoring and accompaniment will 
be carried out on the populations, in the same way the provision of services will be managed 
before the territorial entity of Armenia and the corresponding health entities. of patients' 
health according to the type of insurance. 
Procedure for selecting possible seeds 
Key informants: the first moment will aim to contact key informants who can allow access 
to the seeds; For this purpose, different population groups were approached: for example, 
people in a situation of prostitution, research, care and/or treatment institutions, among other 
agents who are considered key informants during the investigation. 
Seed selection: to access and identify the seeds there will be an experienced team, key 
informants from the institutions or organizations that work with key populations, some natural 
leaders. 
To establish the status of the person as a key population, a small screening questionnaire 
will be designed in which we will seek to establish their status, possible history of 
participation in the study, some socio-demographic data (age, gender, origin, area of 
housing, origin of the network, potential of known people, willingness to also involve them in 
the study). Individuals who are not eligible will be offered information regarding the practice, 
risks, the possibility of referral to an institution for counseling or treatment, and the referral 
potential of other individuals who may meet the inclusion criteria is explored. 
Information collection (expansion phase) 
The person will be summoned to the place where the process will be carried out, upon arrival 
at the site their eligibility is confirmed, if not eligible, the participant will abandon the study, 
otherwise the informed consent will be read, the participant will be registered in the study, 
the structured survey is carried out, then pre-test counseling is provided and the blood 

 
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sample will be taken, performing the rapid test for HCV. The cases will receive the results 
at this same appointment, carrying out post-test counseling. 
Each interview location will have its own screening area in which potential participants will 
be screened before being admitted to the interview area, this will reduce the chances of a 
disgruntled or disruptive person disturbing registered participants or staff. During the 
screening procedure, the person performing the screening will ensure the following: the 
potential participant has an appropriate coupon (correct identification number) and signs the 
informed consent. 
Steps for user detection 
 
Source: self-made 
When the screening steps are completed, the person enrolls in the study and continues with 
the interview. Currently, a registration as a participant is completed. 
In the survey area 
After greeting, the survey is carried out, followed by pre-test counseling (VHC) and the blood 
sample is taken. After completing the survey, the participant is informed about the 
opportunity to recruit voluntarily the importance for the project and the opportunity to receive 
a social support bonus for each eligible person recruited. If the participant is interested, they 
will be indicating the way and how to do it, each participant needs a coupon to participate in 
the study. 
Coupons 
Greet the 
participant
Review voucher 
validity
Locate voucher 
with other 
information
Introduction to the 
screening survey
Ask eligibility 
questions
Verify 
memberships in 
target population
Complete consent 
form
Complete 
participant 
registration

 
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Each of the participants is asked about the number of acquaintances, the coupon system 
limits the possibility for each informant to select only three future informants, to nullify the 
biases caused by the presence of semi-professional recruiters or by a desire for super-
collaboration by some people that would cause an overrepresentation of the networks of a 
specific individual. 
Instructions for recruitment 
• 
They must bring their coupons with them to the site, if the referral does not bring their 
coupon, they cannot participate in the survey. 
• 
Coupons are limited, it is recommended to give the coupons to people you know who 
meet the criteria as soon as possible. 
• 
One hour and 30 minutes are required to complete the interview and sample collection. 
• 
Those referred will receive a social support bonus if they complete the interview and 
sample collection. 
• 
Referrals who are not eligible (not meeting inclusion criteria), participants who do not 
sign the informed consent, who had already participated and if the person sells their 
coupons are not considered (only the social support voucher will be given to the person 
to whom which they were delivered). 
Social network 
They will be consulted about the following aspects corresponding to the social network, 
having the protocol it is expected to have at least the following information: 
• 
How many people do you know (you know them, and they know you)? 
• 
Of them, how many are over 18 years old? 
• 
And of them, how many have you seen in the last month? 
• 
Each person receives three (3) coupons to recruit others. 
• 
The study will end when the required sample size is met. 
• 
A day is scheduled to deliver the social support vouchers. 
Delivery of social support bonuses 

 
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• 
Reiterate the recruitment process to the participant. 
• 
Deliver the primary social support bonus. 
• 
Confirm to the participant that they have completed all steps of the study (otherwise have 
the participant complete them). 
• 
They must bring the coupons, in the part of the detachable assigned for the participants. 
• 
It is investigated whether some people rejected the coupons and the reasons for this 
situation. 
• 
It is consulted for doubts or questions that the participants have. 
• 
Verify that the recruit is enrolled in the study. Inform the recruiter to return if the previous 
coupon was not registered. 
Other activities 
• 
Ensure the quality of the numbers on the coupons. 
• 
Update the coupon registration form. 
• 
Keep the record of primary and secondary compensation updated. 
• 
Match the identification number on the coupon to the participant's recruiter record. 
Coupon Registration 
• 
Record the date of the interview. 
• 
Register the participant's questionnaire code. 
• 
Record the participant's network size. 
• 
Record the identification number of the coupon delivered to the participant. 
• 
Record the identification numbers of the coupons given to the participant to recruit 
others. 
• 
Record important notes or physical features. 
• 
Record the initials of the interviewer. 
 

 
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Post-test advice 
Given the serious ethical implications of developing diagnostic tests, the counseling process 
includes: 
• 
Provide information and education about HCV. 
• 
Referral to prevention, treatment, and support services. The local and departmental 
health secretary has previously been informed about this project for the correct 
identification of the support network and health services. 
• 
Prior to this advisory process, a training session is held for the team responsible for 
collecting information. 
Screening test 
The samples are taken by the personnel assigned to the study with training to ensure the 
quality of the diagnostic process. At the appointment that is made, the rapid test is read to 
deliver the results. 
The sample number will be linked to the survey through a code, which allowed confidentiality 
and anonymity to be maintained without losing the possibility of monitoring the results in 
order to return them to the subjects and offer the corresponding advice and follow-up. . 
Delivery of results with their respective APV 
With the results of the rapid HCV tests, counseling will be carried out after the detection test. 
The goal is to understand the outcome and provide appropriate information, support, and 
reference. If the result is negative, counseling offers an opportunity to promote behaviors 
that reduce infection risks; if the result is positive, to reduce the risk of transmitting the virus 
to other people. In addition, help must be given to face the result and its consequences on 
the person's personal life and that of the person's loved ones. 
Training for interviewers 
The interviewer will be trained in the application of the survey by the technical committee. 
Therefore, each one of them must attend the training to study the definitions and concepts 
well and to become familiar with the procedures and questions of the survey and ask 
questions about any procedure or content that is not clear and so on. Avoid errors as much 
as possible during field work. 

 
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Information processing 
Automated processing will be carried out, the instrument will be designed, tabulated and its 
corresponding database created, tablets will be used with the survey application properly 
adjusted. The first analysis will be carried out in the RDSAT software4, a secondary analysis 
includes other packages such as STATA 10.0, SPSS version 27, with all these programs 
the statistical analysis of the data and the generation of output tables is carried out. 
Results dissemination plan 
At the end of the study, the results are shared and a report with publishable results is 
expected to be generated. 
Social support for respondents in promoting inclusion and diversity 
It is widely known that RDS is a sampling that requires using compensation bonuses that 
motivate the participation of a diverse group of PWIDs, and super-recruiters can be 
controlled and guarantee a close variance between the sample and a population estimate 
without a high homophily; The vouchers also allow you to cover travel expenses from one 
place to another, as well as the time invested to carry out the entire collection and sampling 
procedure. 
As recognition, social support equivalent to $40,000 Colombian pesos will be given for 
participation in the study process cycle and $10,000 Colombian pesos for each recruited 
peer who was eligible and complied with the entire information collection process in the 
study. headquarters, for a maximum of three referrals per participating PWID, will be 
delivered after the administration of the survey and the collection of the blood sample for 
HCV. 
Ethical considerations 
The study protocol will be presented to the institutional human ethics committee of the CES 
University, which is recognized nationally. 
 
4 Respondent Driven Sampling. Available at: http://www.respondentdrivensampling.org/reports/RDSAT60.htm. 
Accessed January 14, 2011. 

 
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The study complies with those established in Resolution 8430 of 1993 of the Ministry of 
Health of Colombia, according to its article 11 numeral b, it is classified as minimal risk, 
given that data records were used through common procedures consisting of a taking a 
blood sample by capillary and/or venous puncture. Furthermore, the criterion of respect for 
the dignity and protection of the rights and well-being of the participants will prevail. 
Written informed consent will be requested from all participants, giving clear and easily 
understandable information about the objectives of the research. Questions about it will be 
explained and resolved, and complete freedom to withdraw consent will be given at any time 
during the study, because the subject considers (Annex 3). 
The study will be coordinated by health professionals and the sampling and collection of 
information will be carried out by trained nursing assistants. The confidentiality of the data 
collected will be guaranteed, the information obtained will not be used for uses other than 
the current research process and at the time of disclosure of the information, no sensitive 
data or data that allows the identification of participating individuals will be published, the 
procedures will be followed. norms set out in law 1581 of 2012 on habeas data. 
The application of rapid tests is governed by current regulations; in particular, by the 
guidelines for carrying out rapid tests outside the clinical laboratory for the early diagnosis 
of hepatitis C infection (Resolution 1314 of 2020 of the Ministry of Health and Social 
Protection). 
In the development of this investigative process, the following ethical aspects will be 
considered by the researchers: 
• 
The integrity of the people interviewed will be respected, regardless of risk behavior, by 
both the interviewers and the researchers. 
• 
Interviewees will be informed of the terms of confidentiality in the informed consent. 
Written consent will be requested from the population to carry out the interview and take 
a blood sample, according to Resolution 8430 of 1993, which regulates research on 
living beings, according to article 14 of this same resolution, consent is understood. 
informed the written agreement, by which the research subject authorizes his or her 
participation in the research, with full knowledge of the nature of the procedures, 
benefits, and risk to which he or she will be subjected, with the capacity of free choice 
and without any coercion. 

 
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• 
In the attention of users, the same procedures will be carried out on all of them; it is 
important to highlight that these instructions will be the same for all users. 
• 
The informed consent that will be signed by all the people who agree to participate in 
the study will clearly explain the justification and objectives of the research, the 
procedures that will be carried out, which will also indicate that there are no relevant 
risks and the benefits. that can be obtained; In addition, the guarantee of receiving an 
answer to any question and clarification of any doubt about the research and other 
matters related to the topic and finally the freedom to withdraw your consent at any time 
and stop participating in the study without thereby create harm in their social 
environment. 
• 
Since the sample is blood, it will be guaranteed to have qualified personnel to take 
samples, as well as guarantee all biosafety procedures and proper handling of blood 
samples. 
• 
The identification number will be linked to the name, telephone number or contact 
information of the person in a database to which only the study coordination and field 
work supervision will have access. 
• 
This study is also governed by the provisions of Chapter IV of Decree 1543/97, which 
establishes: With respect to the right to autonomy, which is none other than self-
determination in the full right of the person's faculties, freely and spontaneous for the 
benefit of the development of their personality, it was very important to include that the 
practice of the diagnostic test for the detection of antibodies against HCV requires the 
informed consent of the person, a requirement without which it cannot be carried out. 
• 
The same Decree establishes the obligation to offer counseling (Article 5). And along 
the same lines, article 6 establishes that the results of tests for diagnosing HCV infection 
and those for diagnosing sexually transmitted diseases (STDs) must be delivered to the 
patient by personnel duly trained in counseling. 
• 
There are suitable personnel in the areas that involve the topics of the study, with 
knowledge and experience to take care of the integrity of the human being to guarantee 
the well-being of the research subject. 
• 
The privacy of the individual will be protected, identifying them only when the results 

 
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require it and they authorize it. 
• 
In relation to confidentiality, Article 34 of Decree 1543/97 establishes that members of 
the health team who know or provide health care to an infected person are obliged to 
keep the consultation, diagnosis, and evolution of the disease confidential. and all the 
information that belongs to your privacy. 
• 
All possible types of risk and the possibility that the research subject will suffer harm as 
an immediate or delayed consequence of the study will be identified. 
• 
Subject participation will be entirely voluntary. 
• 
The cases of other people who have had risky sexual contact and who are not a key 
population of the study, will be notified according to the public health surveillance 
protocol. These actions will be coordinated for this purpose with the territorial entity and 
specifically with the secretaries of health, the latter will be informed from the beginning 
of the project about the scope of the research and the actions that will be carried out; 
positive cases will also be notified within the public health surveillance system of the 
corresponding entity. 
None of the authors have conflicts of interest to declare. 
Expected results and impact 
Strengthening the scientific community 
Expected result/product 
Indicator 
Beneficiary 
Screening for hepatitis C in PWID. Proportion of PWID with a
diagnosis of hepatitis C. 
PWID, users affiliated with 
the SGSSS, benefit plan 
administrative 
entities, 
health personnel. 
Linkage 
to 
the 
treatment 
of 
confirmed positive cases for HCV 
infection at the site included in the 
study. 
Proportion of PWID with a
diagnosis 
of 
chronic 
hepatitis C who started 
treatment. 
PWID, users affiliated with 
the SGSSS, benefit plan 
administrative 
entities, 
health personnel. 
 
 
 

 
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Social appropriation of knowledge 
Expected result/product 
Indicator 
Beneficiary 
Circulation 
of 
specialized 
knowledge: Presentation of results 
at scientific events (national and 
international). 
Participation in scientific 
events presenting research 
results. 
Medical-scientific 
community, 
decision
makers, 
hospital
institutions. 
Presentation 
of 
results 
to 
stakeholders of the Colombian 
SGSSS. 
Participation in an event 
with different stakeholders 
of the SGSSS. 
Insurers 
and 
SGSSS 
providers, 
patients 
nationwide. 
Generation of new knowledge 
Expected result/product 
Indicator 
Beneficiary 
Research article submitted to a 
journal 
indexed 
in 
one 
of 
the
bibliographic citation indexes such as,
for example, ISI-Web of Knowledge
(Science Citation Index [SCI] and
Social Sciences Citation Index [SSCI])
or SCOPUS, Publindex, Lilacs, BVS,
Latin index, among others. 
Published 
scientific
articles. 
Medical-scientific community, 
decision 
makers, 
hospital 
institutions. 
Expected impacts from the use of the results 
Expected impact 
Term (years) 
after project 
completion* 
Verifiable indicator 
Assumptions* 
Improvement in the opportunity 
to detect new cases of hepatitis 
C among PWID. 
1-4 years 
Increase 
in 
the 
incidence 
rate 
for 
hepatitis C. 
Medical 
personnel 
adherent 
to 
the 
screening 
and 
diagnosis algorithm –
applicability 
of 
this 
among PWID. 
Improvement 
in 
the 
times 
elapsed between confirmatory 
diagnosis with PCR and the 
start of treatment. 
1-4 years 
Increase 
in 
the 
number of PWID with 
a diagnosis of chronic 
hepatitis 
C 
who 
started treatment. 
Standardization of the 
route 
that 
allows 
compliance with all 
stages 
for 
the 
diagnosis, treatment,
and 
monitoring 
of 
PWID with HCV. 
Successful 
completion 
of 
treatment between PWID 
1-4 years 
Increase 
in 
the 
proportion of PID with 
cure (sustained viral 
response). 
Opportunity in follow-
up, 
adherence 
of 
PWID to the treatment 
scheme. 

 
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Expected impact 
Term (years) 
after project 
completion* 
Verifiable indicator 
Assumptions* 
Changes in the prevalence 
figures of chronic hepatitis C 
among PWID. 
5-9 years 
Trend 
in 
crude 
prevalence of chronic 
hepatitis C. 
Early detection and 
treatment reduce the
risk of transmission. 
short (1-4), medium (5-9), long (10 or more) 
Schedule of activities 
The activities will be executed in 10 months, the first two months of planning the activities, 
carrying out and submitting the protocol, selecting the field team, two months in the design, 
and execution of the exploratory phase, including the delivery of the unique object; the 
following four months of field activities, and the following two months for the delivery of 
results, and analysis of information, including the reproduction of the final document. 
Activities 
Start date 
Final date 
Development of the final protocol, 
consent reports and approval by 
the ethics committee. 
Month 1 
Month 1 
Nursing 
personal 
training 
for 
information 
capture, 
sample 
collection, referral to treatment 
and follow-up. 
Month 2 
Month 2 
Development phase 1 exploratory 
component. 
Month 3 
Month 4 
Recruitment of participants 
– 
Identification of risk populations to 
be screened – Screening tests – 
entry of captured information. 
Month 5 
Month 7 
Monitoring of positive cases – 
Month 8 
Month 8 

 
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Activities 
Start date 
Final date 
treatment route direction. 
Verification of the quality of the 
information collected. 
Month 5 
Month 8 
Analysis of data. 
Month 9 
Month 9 
Final report presentation. 
Month 10 
Month 10 
Writing and presenting article for 
publication. 
Month 10 
Month 10 
Human resource 
Activities of the city supervisor 
Armenia will have a supervisor, who must guarantee that all the procedures defined for field 
work and completion of the survey are fully complied with. With a professional profile, 
preferably with postgraduate training in the areas of public health, epidemiology and/or 
related. 
Staff activities 
This is the person who is responsible for collecting information in the PWID. To reach the 
sample proposed in the study protocol, it is required that the interviewer be committed to his 
or her work, clearly know the functions he or she must perform, be a responsible, orderly, 
accomplished person, and have certified experience. of work, in addition to having the ability 
to generate empathy and trust, and to demonstrate that they have the basic knowledge 
necessary to contribute to the fulfillment of the objective of the study and to pass the 
evaluation after the training. 
Principal investigator career 
Javier Enrique Hernández Blanco. Medical specialist in Internal Medicine, Gastroenterology 
and Digestive Endoscopy, Master in Liver Diseases and Clinical Epidemiology. University 
Research Professor, president of the Colombian Association of Hepatology (2021-2023). 
Active participant in the working groups of the National plan for the elimination of Viral 

 
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Hepatitis, national lecturer on Viral Hepatitis B and C, former public health advisor to the 
District Health Secretary of Santa Marta, extensive experience in viral hepatitis screening 
activities in the community, co-author of the Colciencias project - CALL 777: Prevalence and 
modeling of Hepatitis C Virus infection in Colombia. 
Budget 
This project will be carried out by the Colombian Association of Hepatology (ACH), it 
corresponds to an initiative of the ACH in response to its commitment to join efforts to 
achieve the World Health Organization's goal of eliminating viral hepatitis by 2030. and is 
financed with resources from the ACH, which in turn are obtained by the association's own 
activities and resources obtained from calls for project financing. Leadership in 
communication and administration of project resources is provided by the ACH. 
A final project report will be generated, and the results will be maintained in a consolidated 
and anonymized manner, without compromising confidential information of the participants. 
The ACH is the author of the project, it will provide the resources for its execution. The items 
covered in the execution of the study do not include the costs of treatment or any clinical 
management procedures (for example, doctor visits, follow-up laboratories) since they are 
part of the usual treatment for hepatitis C virus infection. and it is contemplated in Colombian 
regulations that once the confirmed case of hepatitis C is notified to SIVIGILA (file 340), on 
a quarterly basis the High-Cost Account downloads that information, includes the participant 
in the national cohort of high-cost diseases. Hepatitis C establishes communication/audit 
with the Benefits Plan Administration Company (EAPB) to which the user belongs and 
guarantees their connection to the treatment, treatment against hepatitis C that is free, 
without cost for the EAPB or out-of-pocket expense for the user. user and which is acquired 
in annual purchases by the CAC since 2017 through the centralized purchasing mechanism 
from the Pan American Health Organization. 
 
 

 
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Annexes 
Annex 1 Anti-HCV antibody detection test 
Test principle 
The Bioline™ test5HCV contains a strip with nitrocellulose membrane pre-coated with 
recombinant HCV capture antigen (core, NS3, NS4 and NS5) in the test line area (T). The 
colloidal gold-conjugated protein A and the sample move across the membrane 
chromatographically to the test zone. There, the antigen-antibody complex of protein A 
bound to gold particles forms a visible line with a high level of sensitivity and specificity. On 
the surface, this test device has the letters "T" and "C", which refer to the “test line” and 
“control line”, respectively. The test line and control line are not visible in the results window 
until the sample is applied. The control line is a procedural control. The control line should 
appear if the test procedure is executed correctly, and the control line reagents are 
operational. 
Use 
The Bioline™ HCV Test is a rapid in vitro immunochromatographic assay for the qualitative 
detection of HCV-specific antibodies, in human serum, human plasma (heparin, EDTA and 
sodium citrate) or blood. Bioline™ HCV is exclusively for professional use as an initial test, 
as an aid to diagnosis. Reactive specimens should be reflected for additional testing, either 
using HCV RNA detection technologies or HCV core antigen testing, to identify current HCV 
infection. This product is intended for use in a population with a high prevalence of HCV or 
who has a history of HCV exposure/risk behavior, including pregnant women. This test may 
not be suitable for early diagnosis of infection or for testing blood donation. 
Materials included and active ingredients of the main components 
1. 25 test devices with desiccant in individual aluminum foil bags. 
2. Assay diluent (1 x 5 ml/vial). 
3. 25 capillary pipettes (10 μl), 25 safety lancets, 25 alcohol swabs. 
 
5Anti-HCV antibody detection test. Reference 02FK10/02FK16/02FK17. Abbott Diagnostics Korea Inc. Date 
issued: 2020. 04. 

 
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Materials needed, but not provided in the kit: Micropipette, protective gloves, timer, container 
for biological waste. 
Kit Storage and Stability 
1. The test kit should be stored at a temperature between 1°C and 30°C. Do not freeze the 
kit or its components. 
2. The assay diluent can be opened and resealed for each assay. The lid should be tightly 
sealed between each use. The assay diluent is stable until the expiration date if kept at 1 
and 30 °C. 
3. The test device is sensitive to heat and humidity. Perform the test immediately after 
removing the test device from the aluminum foil bag. 4. Do not use the test kit after the 
expiration date. The shelf life of the kit is indicated on the outer packaging. 
5. Do not use the test kit if the bag is damaged or the seal has been breached. 
Warnings 
1. Test devices are for in vitro diagnostic use only. Do not reuse the test device. 
2. Follow the instructions properly to get accurate results. The person in charge of testing 
this product must be a specialist trained to use it. 
3. Do not mix or exchange different samples. 
4. Do not put the desiccant in the aluminum foil bag in your mouth. 
5. Do not mix or interchange components from different lots or other products. 
6. Take care to avoid contamination of the container spout when pouring the assay diluent 
into the sample well. 
Sample collection and storage 
1. Blood 
[Venipuncture collection] 
• 
Perform a venipuncture and place the blood in the collection tube (containing 
anticoagulants, such as heparin, EDTA, and sodium citrate). 
• 
If the test is not performed immediately on the blood sample, it should be refrigerated at 

 
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2°C to 8°C. 
• 
If the blood sample is stored between 2°C and 8°C, the test should be performed within 
3 days of refrigeration. 
• 
Do not use blood samples stored for more than 3 days, as a non-specific reaction may 
occur. 
• 
Before testing, the blood sample should be at room temperature (between 15°C and 
30°C). 
[Lancet collection] 
• 
Clean the area where you will use the lancet with an alcohol swab. 
• 
Squeeze the tip of your finger and then prick the side of your finger with the lancet 
provided. Wipe away the first drop of blood. Immediately afterwards, dispose of the 
lancet safely. 
• 
Dip the open end of a new capillary pipette (10 μL) into the next drop of blood and relieve 
the pressure to draw blood into the capillary pipette until the fill line is reached. 
Plasma or serum 
[Plasma] Using venipuncture, introduce the blood into the collection tube (containing 
anticoagulants, such as heparin, EDTA, and sodium citrate), then centrifuge to obtain the 
plasma sample. 
[Serum] Using venipuncture, introduce the blood into the collection tube (which DOES NOT 
contain anticoagulants) and let it rest for 30 minutes for coagulation to occur. Centrifuge the 
tube to obtain a serum sample. 
If plasma or serum samples will not be used immediately, they should be refrigerated at 2°C 
to 8°C. If it is necessary to store them for more than 2 weeks, they must be frozen (Less 
than -20 °C). Before testing, the plasma or serum sample should be at room temperature 
(between 15°C and 30°C). 
Plasma or serum samples containing precipitate may give inconsistent test results. It is 
necessary to clarify the samples by centrifugation before testing. 
 
 

 
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Test procedure 
1. Before testing, bring all kit components and samples to a room temperature between 15°C 
and 30°C. 
2. Remove the test device from the bag and place it on a flat, dry surface. Label the test 
device with a patient identifier. 
3. [Using a micropipette] Place 10 µl of the plasma, serum or blood sample into the sample 
well marked “S”. Or [Using a capillary pipette] Place 10 µl of the drawn blood sample into 
the sample well marked “S”. 
 
4. Place 4 drops of assay diluent into the “S” sample wells. 
Caution: If the bottle is not held vertically, inadequate results may be obtained. Add exactly 
4 drops. Adding more than 4 drops may cause the background to turn reddish or cause an 
erroneous result. Do not allow the tip of the bottle to come into contact with the device to 
avoid cross contamination. 

 
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5. When the test starts running, you will see a purple color scrolling across the results 
window located in the center of the test device. 
6. 5 to 20 minutes after adding the assay diluent, interpret the results. Do not read the results 
after 20 minutes. 
 
Caution: If the test result is not readable after 5 minutes due to the intense color of the 
background, read it again later, within 20 minutes of adding the diluent. Reading the result 
outside of that time frame (before 5 minutes or after 20 minutes) may give false results. 
Test interpretation 
1. A control line will appear in the "C" section of the results window indicating that the test is 
working correctly. 2. In the "T" section of the results window, the test results are displayed. 

 
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Non-reactive result: If only the control line (C) appears in the result window, the result is non-
reactive. 
 
Reactive Result: If the test line (T) and the control line (C) appear in the result window, 
regardless of the order of appearance, the result is reactive. Caution: The presence of any 
test line, even a faint color, indicates that the result is reactive. 
 
Invalid result: If the control line (C) is not visible in the results window after running the test, 
it is considered that there is no valid result. This situation may be because the instructions 
were not followed correctly or because the test has deteriorated. It is recommended that the 
sample be retested with a new test device. 
 
Internal quality control 
The Bioline™ HCV Test Device has two pre-coated lines on the test surface: “T” (test line) 
and “C” (control line). The test line and control line are not visible in the results window 

 
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before sample application. The control line is used as a reference in the procedure and only 
indicates that the diluent was applied correctly and that the active ingredients of the main 
components of the test strip work but does not ensure correct application of the sample; It 
is not a reactive sample control. 
Performance characteristics 
The Bioline™ HCV Test Kit is designed to have 99.3% (95% CI: 96.1 - 99.9%) sensitivity 
and 98.1% (95% CI: 94.5 - 99.4%) sensitivity. %) specificity. 
References 
1.  
World Health Organization (WHO). Hepatitis-C [Internet]. 2020. Available from: 
http://www.who.int/es/news-room/fact-sheets/detail/hepatitis-c 
2.  
Taherkhani R, Farshadpour F. Global elimination of hepatitis C virus infection: 
Progresses and the remaining  challenges. World J Hepatol. 2017 Nov;9(33):1239–
52.  
3.  
World Health Organization (WHO). Guidelines for the care and treatment of persons 
diagnosed with chronic hepatitis C virus infection [Internet]. Available from: 
https://www.who.int/publications/i/item/9789241550345 
4.  
Fuller CM, Ompad DC, Galea S, Wu Y, Koblin B, Vlahov D. Hepatitis C incidence--a 
comparison between injection and noninjection drug users  in New York City. J Urban 
Health. 2004 Mar;81(1):20–4.  
5.  
Nelson PK, Mathers BM, Cowie B, Hagan H, Des Jarlais D, Horyniak D, et al. Global 
epidemiology of hepatitis B and hepatitis C in people who inject drugs:  results of 
systematic reviews. Lancet (London, England). 2011 Aug;378(9791):571–83.  
6.  
Keen L 2nd, Khan M, Clifford L, Harrell PT, Latimer WW. Injection and non-injection 
drug use and infectious disease in Baltimore City:  differences by race. Addict Behav. 
2014 Sep;39(9):1325–8.  
7.  
World Health Organization (WHO). People who inject drugs [Internet]. Available from: 
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