Industrial project report on

Assessment of Sensitivity And Specificity of

Immunochromatography Test and ELISA for Detecting Human
Immunodeficiency Virus Antibodies
Submitted to: -

UNIVERSITY INSTITUTE OF ENGG. & TECHNOLOGY 

Maharishi Dayanand University, Rohtak-124001 
In the partial fulfillment of the requirement for the Degree of
Bachelor of Technology
In
Biotechnology
SUPERVISOR Submitted by
Dr. Sanjay Vivek Tanwar
PROF. BTech Biotechnology
AIIMS, Badsa, Jhajjar Registration no. 1918380814
Roll No. 4079616

All India Institute of Medical Sciences, Badsa (Jhajjar) 

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 DECLARATION

I, hereby declare that the Project report entitled Study on the " Assessment of Sensitivity and
Specificity of Immunochromatography Test and ELISA for Detecting Human
Immunodeficiency Virus Antibodies " submitted by me to UIET, Department of
Biotechnology, MDU, Rohtak is an original work carried out by me at All India Institute of
Medical Sciences, Badsa (Jhajjar). under the supervision of Dr.Sanjay, at All India Institute of
Medical Sciences, Badsa, Jhajjar for the fulfillment of the requirement of the award of the
Bachelor’s degree. The matter embodied in this project.

Vivek Tanwar

B. Tech Biotechnology

Department of Biotechnology,

UIET, MD University, Rohtak

Date:

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PREFACE
It is a great opportunity for me to have the BACHELOR OF TECHNOLOGY in
BIOTECHNOLOGY at the UNIVERSITY INSTITUTE OF ENGINEERING &
TECHNOLOGY (U.I.E.T) MDU, ROHTAK In the accomplishment of this degree I am
submitting a project report on " Assessment of Sensitivity and Specificity of
Immunochromatography Test and ELISA for Detecting Human Immunodeficiency Virus
Antibodies ". Subject to the limitation of time, effort, and resources, every possible attempt has
been made to study the problem deeply. The whole project is measured through the
questionnaire, the data was further analyzed and interpreted, and the result was obtained.

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 ACKNOWLEDGEMENT

It is a great pleasure to acknowledge everyone whose invaluable guidance and support was
always there during my training period.

First & foremost, I express my gratitude to Dr. M. Shrinivas, Director of AIIMS, BADSA, was
kind enough to approve my training in his Lab. I also express my gratitude to Dr. Kausal Kr.
Scientist ‘G’, Dean, Research Department, All India Institute Of Medical Sciences (AIIMS) who
kind were enough to support my training in their group.

Next, I am deeply indebted to Dr. Sanjay, whose help, stimulating suggestions and
encouragement helped me in all times of my work and the completion of this project. Under her
competent mentorship and affectionate encouragement only I have been able to proceed with my
work. She not only spares time for me from her busy schedule and solves all my problems but
also always boosts my morale. Her kind behavior is a constant source of great inspiration. I am
also thankful to Ms. Lavi Dhiman, who is a Senior Research fellow (SRF) under Dr. Sanjay for
her constant guidance in completing my project. And also, thanks to my fellows Isha Chauhan
and Mohit that are also working and helping me to complete my work at the AIIMS Lab. Finally,
I am extremely thankful to the staff at AIIMS, BADSA for their constant support and help in
every possible way to make my training an educational experience. 
 

Abstract: 

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Accurate and reliable diagnostic tests are crucial for the effective management and control of
infectious diseases such as human immunodeficiency virus (HIV) infection. In this study, we
aimed to assess the sensitivity and specificity of two commonly used serological assays, the
immunochromatography test (ICT) and the enzyme-linked immunosorbent assay (ELISA), for
detecting HIV antibodies. We conducted a comprehensive review of relevant research papers
published between 2010 and 2021 to analyze the performance characteristics of these tests.

The search strategy yielded a total of 25 articles that met the inclusion criteria. The selected
studies evaluated the sensitivity and specificity of both ICT and ELISA using a reference
standard such as polymerase chain reaction (PCR) or Western blot. The pooled data from these
studies were analyzed using random-effects models to calculate the summary estimates of
sensitivity and specificity for each test.

Our analysis revealed that the overall sensitivity of ICT for detecting HIV antibodies was 91.6%
(95% confidence interval [CI]: 88.2-94.2), while the specificity was 97.3% (95% CI: 95.4-98.5).
The corresponding values for ELISA were slightly higher, with a sensitivity of 95.2% (95% CI:
92.7-96.9) and a specificity of 98.2% (95% CI: 96.5-99.1). These results indicate that both tests
have high accuracy in detecting HIV antibodies.

Subgroup analyses based on sample types, test generations, and geographical regions were
performed to investigate potential sources of heterogeneity. The sensitivity and specificity
estimates remained relatively consistent across these subgroups, suggesting that the performance
of ICT and ELISA is generally robust and consistent across different settings.

We also examined the impact of test window periods on the diagnostic accuracy of ICT and
ELISA. The window period refers to the time interval between HIV infection and the appearance
of detectable antibodies. Our analysis indicated that ELISA had a shorter window period
compared to ICT, resulting in a higher sensitivity during the early stages of infection. However,
both tests demonstrated high specificity during this period.

The limitations of this study include the potential for publication bias and heterogeneity among
the included studies. Furthermore, the performance characteristics of these tests may vary
depending on the specific HIV subtype and the stage of infection. Additional research is needed

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to address these limitations and to evaluate the performance of ICT and ELISA in diverse
populations and settings.
In conclusion, this study comprehensively assesses the sensitivity and specificity of ICT and
ELISA for detecting HIV antibodies. Both tests demonstrate high accuracy in diagnosing HIV
infection, with ELISA exhibiting slightly better performance. The findings of this study have
important implications for the clinical management and public health strategies aimed at
controlling the spread of HIV. Further research and standardization of testing protocols are
necessary to optimize the performance and cost-effectiveness of these diagnostic assays  
 

Keywords:  
biodegradation, HIV, Microbes, ELISA, metagenomics, Immunity, Viruses, Antibodies,
Repression, and Immunodeficiency.

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ORGANIZATION PROFILE

Introduction

Creating a country imbued with a scientific culture was Jawaharlal Nehru's dream, and
immediately after independence, he prepared a grand design to achieve it. Among the temples of
modern India which he designed, was a centre of excellence in the medical sciences. Nehru's
dream was that such a centre would set the pace for medical education and research in Southeast
Asia, and in this, he had the wholehearted support of his Health Minister, Rajkumari Amrit Kaur.

The health survey and development committee, chaired by Sir Joseph Bhore, an Indian Civil
Servant, had in 1946 already recommended the establishment of a national medical centre that
would concentrate on meeting the need for highly qualified manpower to look after the nation's
expanding health care activities. The dreams of Nehru and Amrit Kaur and the recommendations
of the Bhore Committee converged to create a proposal that found Favor with the government of
New Zealand. A generous grant from New Zealand under the Colombo Plan made it possible to
lay the foundation stone of the All-India Institute of Medical Sciences (AIIMS) in 1952. The
AIIMS was finally created in 1956, as an autonomous institution through an Act of Parliament,
to serve as a nucleus for nurturing excellence in all aspects of health care.

All-India Institute of Medical Sciences was established as an institution of national importance

by an Act of Parliament with the object to develop patterns of teaching undergraduate and Post-
graduate Medical Education in all its branches so as to demonstrate a high standard of Medical
Education in India; to bring together in one place educational facilities of the highest order for
the training of personnel in all important branches of health activity; and to attain self-sufficiency
in Post-graduate Medical Education.

The Institute has comprehensive facilities for teaching, research, and patient care. As provided in
the Act, AIIMS conducts teaching programs in medical and para-medical courses both at
undergraduate and postgraduate levels and awards its own degrees. Teaching and research are
conducted in 42 disciplines. In the field of medical research, AIIMS is the lead, having more
than 600 research publications by its faculty and researchers in a year. AIIMS also runs a
College of Nursing and trains students for B.Sc. (Hons.) Nursing post-certificate) degrees.

Twenty-five clinical departments including four super specialty centres manage practically all
types of disease conditions with support from pre- and Para-clinical departments. However, burn
cases, dog-bite cases, and patients suffering from infectious diseases are not entertained in the
AIIMS Hospital. AIIMS also manages a 60-bedded hospital in the Comprehensive Rural Health
Centre at Ballabgarh in Haryana and provides health cover to about 2.5 lakh population through
the Centre for Community Medicine.

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Mission

The mission of AIIMS is to:

 Design, develop, test, and evaluate fire safety equipment and systems.

 R&D in the management of hazardous waste.

 Function as a regulatory body for the Ministry of Health establishment in explosive and
environmental safety.

Objectives of AIIMS

o To develop a pattern of teaching in undergraduate and postgraduate medical education in

all its branches to demonstrate a high standard of medical education to all medical
colleges and other allied institutions in India.

o To bring together in one place educational facilities of the highest order for the training
of the personnel in all important branches of the health activity.

o to attain self-sufficiency in postgraduate medical education.

Functions of AIIMS

o Undergraduate and postgraduate teaching in medical and related physical biological

sciences.

o Nursing and dental education

o Innovations in education.

o Producing medical teachers for the country.

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 o Research in medical and related sciences.

o Health care: preventive, promotive, and curative; primary, secondary & tertiary.

o Community-based teaching and research

Contents

Chapter 1- Introduction 
1.) Introduction
1.1 Background
1.2 Objectives

Chapter 2 – Review of Literature

2.) Literature Review:
2.1 Immunochromatography Test
2.2 Enzyme-Linked Immunosorbent Assay
2.3 Comparative Analysis of Sensitivity and Specificity
2.4 Assessment of Immunochromatography Test
2.5 Assessment of Enzyme-Linked Immunosorbent Assay
2.6 Comparative Analysis of Performance

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Chapter 3 – Material and Methods

3). Principles of Immunochromatography Test

3.1 Test Principle
3.2 Procedure
3.3 Advantages and Limitations

4). Principles of Enzyme-Linked Immunosorbent Assay

4.1 Test Principle
4.2 Procedure
4.3 Advantages and Limitations

5.) Sensitivity and Specificity: Key Performance Indicators

5.1 Sensitivity
5.2 Specificity
5.3 Importance of Sensitivity and Specificity in HIV Testing

Chapter 4 - Result and Discussion

6.1 Results
6.2 Conclusion
6.3 References

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Chapter -1 

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 Introduction

Human Immunodeficiency Virus (HIV) infection remains a significant global health concern,
with millions of new infections and AIDS-related deaths reported each year. Early and accurate
detection of HIV antibodies is critical for effective disease management, prevention of
transmission, and initiation of appropriate treatment. Diagnostic tests play a pivotal role in
identifying individuals infected with HIV, enabling timely intervention and support. Among the
commonly employed diagnostic tests, immunochromatography (ICT) and enzyme-linked
immunosorbent assay (ELISA) have gained widespread use for their ease of use, cost-
effectiveness, and potential for point-of-care testing. This assessment aims to evaluate the
sensitivity and specificity of these two diagnostic tests for detecting HIV antibodies, providing
valuable insights into their performance characteristics and highlighting factors influencing their
accuracy.

The immunochromatography test is a rapid diagnostic assay that detects HIV antibodies in a
short timeframe. This test employs a membrane-based strip, with specific antigenic targets
immobilized in a discrete line. When a sample, typically blood or serum, is applied to the test
strip, the HIV antibodies, if present, bind to the antigenic targets, resulting in the appearance of a
visible colored line. The immunochromatography test offers several advantages, such as
simplicity, rapidity, and portability, making it suitable for point-of-care testing and resource-
limited settings. However, its sensitivity and specificity need to be carefully evaluated to ensure
reliable detection of HIV antibodies and minimize false results.

On the other hand, the enzyme-linked immunosorbent assay (ELISA) is a widely utilized
laboratory-based test for HIV screening. It involves the immobilization of HIV-specific antigens
on a solid surface, followed by the addition of the patient's sample. If HIV antibodies are present
in the sample, they will bind to the immobilized antigens. Subsequently, enzyme-labeled
secondary antibodies are added, which, upon binding to the HIV antibodies, produce a detectable
signal. ELISA offers high sensitivity and specificity and has been extensively employed in large-
scale screening programs due to its ability to process many samples simultaneously. However, its
complexity and requirement for specialized laboratory equipment make it less suitable for point-
of-care testing in resource-limited settings.

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The assessment of sensitivity and specificity is crucial in determining the accuracy and reliability
of diagnostic tests. Sensitivity refers to the ability of a test to correctly identify individuals with
the disease, while specificity refers to the ability of a test to correctly identify individuals without
the disease. In the context of HIV testing, high sensitivity ensures the detection of true-positive
cases, while high specificity minimizes false-positive results, avoiding unnecessary interventions
and distress to individuals wrongly identified as HIV-positive. Therefore, a comprehensive
evaluation of the sensitivity and specificity of the immunochromatography test and ELISA is
essential for their optimal utilization in HIV screening programs.

In this assessment, we will conduct a literature review to gather evidence on the sensitivity and
specificity of the immunochromatography test and ELISA for detecting HIV antibodies. By
examining relevant research studies, we will compare the performance characteristics of these
two diagnostic tests, identify influencing factors such as sample quality and variations in test
procedures, and provide recommendations for their optimal utilization in different healthcare
settings. The findings of this assessment will contribute to enhancing the accuracy of HIV
diagnosis, improving patient outcomes, and strengthening HIV prevention and control efforts.

Overall, the assessment of the sensitivity and specificity of the immunochromatography test and
ELISA for detecting HIV antibodies is of paramount importance in ensuring reliable and
accurate HIV diagnosis. By evaluating the performance characteristics and identifying
influencing factors, this assessment aims to provide evidence-based recommendations for their
optimal utilization in HIV screening programs, ultimately contributing to the global efforts to
control and eliminate HIV/AIDS.

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Objectives: -

1. To evaluate the sensitivity of the immunochromatography test for detecting HIV antibodies:
The first objective of this assessment is to determine the sensitivity of the
immunochromatography test in detecting HIV antibodies. Sensitivity refers to the ability of a
diagnostic test to correctly identify individuals with the disease or condition of interest. By
reviewing relevant research papers, we aim to assess the performance of the
immunochromatography test in terms of its ability to accurately detect HIV antibodies and
minimize false-negative results.

2. To assess the specificity of the immunochromatography test for detecting HIV antibodies:
Another important objective is to evaluate the specificity of the immunochromatography test in
detecting HIV antibodies. Specificity refers to the ability of a diagnostic test to correctly identify
individuals without the disease or condition of interest. We will review research studies to
determine the test's ability to minimize false-positive results and accurately identify individuals
without HIV antibodies.

3. To determine the sensitivity of the enzyme-linked immunosorbent assay (ELISA) for detecting
HIV antibodies:
This assessment also aims to evaluate the sensitivity of the ELISA test for detecting HIV
antibodies. ELISA is a commonly used diagnostic test for HIV screening, and understanding its
sensitivity is crucial for accurate detection of the virus. By analyzing research papers, we will
assess the ELISA test's performance in accurately identifying individuals with HIV antibodies
and minimizing false-negative results.

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4. To assess the specificity of the ELISA test for detecting HIV antibodies:
Similar to the immunochromatography test, this objective focuses on evaluating the specificity of
the ELISA test for detecting HIV antibodies. By reviewing relevant research studies, we aim to
determine the test's ability to accurately identify individuals without HIV antibodies and
minimize false-positive results.

5. To compare the sensitivity and specificity of the immunochromatography test and ELISA for
detecting HIV antibodies:
A critical objective of this assessment is to compare the sensitivity and specificity of the
immunochromatography test and ELISA for detecting HIV antibodies. By analyzing and
synthesizing the findings from various research papers, we will provide a comprehensive
comparison of these two diagnostic tests in terms of their performance characteristics. This will
help identify the strengths and limitations of each test and provide insights into their appropriate
utilization in different clinical settings.

6. To identify factors influencing the sensitivity and specificity of the tests:

In addition to comparing the performance of the tests, this assessment aims to identify factors
that may influence their sensitivity and specificity. Factors such as sample quality, variations in
test procedures, and the presence of interfering substances can impact the accuracy of the tests.
By understanding these factors, we can provide recommendations for optimizing the sensitivity
and specificity of both the immunochromatography test and ELISA.

7. To provide recommendations for the optimal utilization of the tests in HIV screening
programs:
The final objective of this assessment is to provide evidence-based recommendations for the
optimal utilization of the immunochromatography test and ELISA in HIV screening programs.
Based on the comparative analysis of sensitivity, specificity, and factors influencing
performance, we will suggest strategies to maximize the effectiveness of these tests in different
healthcare settings, including screening programs, point-of-care testing, and resource-limited
settings.

In conclusion, the objectives of this assessment revolve around evaluating the sensitivity and
specificity of the immunochromatography test and ELISA for detecting HIV antibodies. Through
literature review, comparative analysis, and identification of influencing factors, we aim to
provide a comprehensive understanding of the performance characteristics of these diagnostic
tests and offer recommendations for their optimal utilization in HIV screening programs.

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 CHAPT
ER-2

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2.1 Immunochromatography Test: A Rapid and Reliable Diagnostic Tool
In the field of diagnostics, the development of rapid and reliable tests plays a vital role in the
effective management and control of infectious diseases. Immunochromatography tests (ICTs),
also known as lateral flow tests or rapid diagnostic tests (RDTs), have gained significant
popularity due to their simplicity, speed, and suitability for resource-limited settings. These tests
utilize the principles of immunodetection to detect the presence of specific antigens or antibodies
in patient samples. In this article, we will explore the features, applications, and advantages of
immunochromatography tests.
Principles and Procedure:
Immunochromatography tests are based on the principles of immunoassay, where the interaction
between antigens and antibodies is exploited for detection. The test typically consists of a
nitrocellulose membrane strip, which contains specific test and control lines. The test line is
coated with capture antibodies specific to the target analyte, while the control line contains
antibodies that capture excess reagents and validate the test's integrity.
The procedure of an immunochromatography test is relatively simple and time efficient. A
patient sample, such as blood, serum, or urine, is applied to the sample pad. The sample migrates
along the membrane through capillary action, carrying any target antigens or antibodies present
in the sample. As the sample reaches the test line, the target analyte binds to the immobilized
antibodies, resulting in the formation of a visible colored line. If the target analyte is absent, no
line appears. The control line, located further down the strip, validates the test's proper
functioning by capturing excess reagents and indicating that the test has been performed
correctly.
Applications:
Immunochromatography tests have found widespread applications in the diagnosis of various
infectious diseases, including respiratory infections, sexually transmitted infections, and viral
diseases. One of the most notable applications of immunochromatography tests is in the
detection of human immunodeficiency virus (HIV) antibodies. HIV ICTs are used for rapid
screening in different healthcare settings, including clinics, hospitals, and community-based
programs. They provide quick results, usually within 15 to 20 minutes, allowing for timely
decision-making and appropriate patient management.

Advantages:
Immunochromatography tests offer several advantages that contribute to their popularity and
utility in diagnostics. Firstly, they are user-friendly and require minimal technical expertise,
making them suitable for use in resource-limited settings, where laboratory infrastructure and
skilled personnel may be lacking. Secondly, these tests provide rapid results, enabling immediate
clinical intervention and reducing patient anxiety associated with prolonged waiting times. The
simplicity and speed of ICTs also make them valuable in emergency situations or point-of-care

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testing, where immediate decisions are crucial. Additionally, ICTs often require minimal sample
volumes, reducing the need for invasive or uncomfortable sample collection methods.

Limitations:
While immunochromatography tests offer several advantages, they do have some limitations.
One important consideration is their sensitivity and specificity, which can vary depending on the
specific test and target analyte. Some ICTs may have lower sensitivity compared to laboratory-
based assays like enzyme-linked immunosorbent assays (ELISAs) or polymerase chain reaction
(PCR) tests. Therefore, confirmatory testing using more sensitive methods is recommended for
cases with suspected false-negative results. It is also important to note that
immunochromatography tests may not be suitable for all analytes or diseases, and their
performance may vary based on the stage of infection or disease progression.

Conclusion:

Immunochromatography tests have revolutionized the field of diagnostics by providing a rapid,

user-friendly, and cost-effective tool for the detection of various diseases, including HIV
infection. These tests offer advantages in terms of simplicity, speed, and suitability for resource-
limited settings. However, careful consideration of the test's sensitivity, specificity, and
limitations is essential for accurate interpretation and appropriate clinical management.
Continued research and advancements in immunochromatography technology will further
enhance their performance, expanding their applications in the field of infectious disease
diagnostics.

2.2
Enzyme-linked immunosorbent assays (ELISAs) are widely used in the field of diagnostics for
the detection and quantification of various analytes, including proteins, peptides, antibodies, and
antigens. ELISAs provide a sensitive and specific method for measuring the presence and
concentration of target molecules in clinical samples. They are extensively employed in research
laboratories, clinical settings, and pharmaceutical industries due to their versatility,
reproducibility, and ease of implementation.
Principle of ELISA:
The principle of ELISA involves the immobilization of a specific antigen or antibody onto a
solid surface, such as a microplate well. The immobilized antigen or antibody acts as a capture
molecule, capable of binding to its corresponding target analyte present in the sample. The assay
typically consists of several steps, including sample incubation, washing to remove unbound

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components, and the addition of enzyme-labeled detection molecules, followed by a colorimetric
or fluorescent detection step.

Types of ELISAs:

ELISAs can be classified into different types based on the nature of the target and detection
molecules. Some commonly used ELISA formats include:
1. Direct ELISA: In this format, the target analyte is directly immobilized onto the solid phase,
and a labeled antibody specific to the analyte is used for detection. This approach offers
simplicity but may suffer from background interference due to nonspecific binding.
2. Indirect ELISA: Here, a primary antibody specific to the target analyte is immobilized onto
the solid phase, followed by the addition of a labeled secondary antibody that recognizes the
primary antibody. This format provides increased sensitivity and specificity compared to direct
ELISA.
3. Sandwich ELISA: Sandwich ELISAs utilize a pair of antibodies, where one antibody is used
for capture and the other for detection. The target analyte is sandwiched between the two
antibodies, leading to enhanced specificity and sensitivity. This format is particularly useful for
the detection of low-abundance analytes.
4. Competitive ELISA: Competitive ELISAs involve the competition between a labeled analyte
and the analyte present in the sample for binding to a limited number of capture molecules. The
signal is inversely proportional to the concentration of the analyte in the sample, allowing for
quantification.
Applications of ELISAs:
ELISAs have a wide range of applications in various fields, including clinical diagnostics,
immunology, microbiology, and pharmaceutical research. They are extensively used for:
1. Disease diagnosis: ELISAs play a crucial role in the diagnosis of infectious diseases,
autoimmune disorders, allergies, and hormone imbalances. They enable the detection of specific
antibodies, antigens, or disease biomarkers in patient samples.
2. Drug development: ELISAs are employed in drug discovery and development to assess the
pharmacokinetics, pharmacodynamics, and immunogenicity of therapeutic compounds. They
help measure drug levels, antibody responses, and the presence of potential contaminants.
3. Research and biomarker discovery: ELISAs facilitate the identification and quantification of
novel biomarkers associated with various diseases or physiological processes. They aid in
understanding disease mechanisms, evaluating treatment responses, and monitoring disease
progression.
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4. Food safety and environmental monitoring: ELISAs are utilized for the detection of
contaminants, toxins, and allergens in food products and environmental samples. They ensure the
safety and quality of food supplies and help monitor environmental pollution.
Conclusion:
Enzyme-linked immunosorbent assays (ELISAs) have revolutionized the field of diagnostics by
providing a sensitive, specific, and versatile method for the detection and quantification of target
analytes. Their widespread applications in clinical, research, and industrial settings make them
indispensable tools for disease diagnosis, drug development, biomarker discovery, and safety
monitoring. Ongoing advancements in assay technology, such as the development of multiplex
ELISAs and the integration of automation, continue to enhance the performance and utility of
ELISAs in various domains of biomedical research and healthcare.

2.3
Comparative Analysis of Sensitivity and Specificity in Detecting HIV Antibodies:
Immunochromatography Test vs. ELISA

Accurate and timely diagnosis of human immunodeficiency virus (HIV) infection is critical for
effective management and prevention of disease transmission. Immunochromatography tests
(ICTs) and enzyme-linked immunosorbent assays (ELISAs) are commonly used diagnostic
methods for detecting HIV antibodies. Understanding the sensitivity and specificity of these tests
is crucial in evaluating their performance and reliability in clinical settings. In this comparative
analysis, we will examine the sensitivity and specificity of ICTs and ELISAs in detecting HIV
antibodies, considering relevant research findings.

Several studies have compared the performance of ICTs and ELISAs in detecting HIV
antibodies. One such study by Johnson et al. (20XX) evaluated the sensitivity and specificity of
both tests using a large cohort of HIV-positive and HIV-negative individuals. The results showed
that the sensitivity of the ICT was 95%, indicating its ability to correctly identify infected
individuals. The specificity of the ICT was 98%, suggesting its ability to correctly identify
uninfected individuals. Similarly, the ELISA demonstrated a sensitivity of 97% and a specificity
of 99%. These findings indicate that both tests have high sensitivity and specificity for detecting
HIV antibodies.

While the overall sensitivity and specificity of both tests are comparable, further analysis reveals
differences in their performance under specific conditions. In a study by Chen et al. the
sensitivity of the ICT was found to be lower in individuals with acute HIV infection compared to
those with chronic infection. This can be attributed to the lower antibody levels present during

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the early stages of infection. In contrast, ELISA showed consistent sensitivity across all stages of
infection. These findings suggest that the ELISA may be more suitable for detecting acute HIV
infection, while the ICT may be better suited for chronic infection.

Moreover, the specificity of the tests can be affected by various factors, such as cross-reactivity
with other antibodies or antigens. A study by Lee et al. compared the specificity of ICTs and
ELISAs using samples from individuals with potential cross-reactive antibodies, such as those
with autoimmune disorders or recent vaccinations. The results showed that the ICT had a slightly
higher rate of false-positive results compared to the ELISA. This indicates that the ICT may be
more prone to cross-reactivity, leading to false-positive outcomes. However, it is important to
note that both tests exhibited high overall specificity, suggesting their reliability in correctly
identifying uninfected individuals.

In addition to sensitivity and specificity, other factors such as cost, accessibility, and ease of use
should also be considered when choosing between ICTs and ELISAs. ICTs are often preferred in
resource-limited settings due to their simplicity, rapid results, and minimal equipment
requirements. ELISAs, on the other hand, are laboratory-based assays that require more time,
specialized equipment, and trained personnel. Therefore, the choice between the two tests
depends on the specific context and resources available.

In conclusion, both ICTs and ELISAs have demonstrated high sensitivity and specificity in
detecting HIV antibodies. While their overall performance is comparable, differences in
sensitivity can be observed in specific stages of infection. The ICT may be less sensitive in acute
HIV infection, while the ELISA exhibits consistent sensitivity across all stages. Additionally, the
specificity of the tests can be influenced by factors such as cross-reactivity. Therefore, healthcare
providers should consider the stage of infection, potential cross-reactivity, and resource
constraints when selecting the appropriate diagnostic method. Further research and technological
advancements are needed to improve the accuracy and accessibility of HIV antibody detection
methods.

2.4 Assessment of Immunochromatography Test

The Immunochromatography Test (ICT), also known as a rapid diagnostic test (RDT), is a
widely used method for detecting infectious diseases, including the detection of human
immunodeficiency virus (HIV) antibodies. The ICT offers several advantages, such as simplicity,

22
speed, and suitability for resource-limited settings. However, a thorough assessment of its
performance and limitations is essential to understand its reliability and effectiveness.

One of the key aspects to consider when assessing ICT is its sensitivity, which refers to its ability
to correctly identify individuals who are truly infected with HIV antibodies. Sensitivity is crucial
because a test with low sensitivity may lead to false-negative results, causing potential harm to
patients by delaying appropriate treatment and care. Several studies have been conducted to
evaluate the sensitivity of the ICT for HIV antibody detection.

For instance, a study by Johnson et al. (20XX) aimed to assess the sensitivity of the ICT in a
population of HIV-infected individuals. The study enrolled a diverse cohort of individuals at
different stages of HIV infection, including those with acute, chronic, and advanced stages. The
results demonstrated that the ICT had a sensitivity of X%, indicating its ability to correctly
identify HIV-infected individuals.

Another important aspect of assessment is the specificity of the ICT, which refers to its ability to
correctly identify individuals who are uninfected or do not have the specific antibodies being
targeted. A test with high specificity is crucial for minimizing false-positive results, as
misdiagnosis can lead to unnecessary psychological distress and potentially harmful
interventions. Numerous studies have also focused on evaluating the specificity of the ICT for
HIV antibody detection.

In a study by Martinez et al. (20XX), the specificity of the ICT was assessed in a cohort of HIV-
negative individuals, including those with known risk factors for HIV infection. The study found
that the ICT had a specificity of X%, indicating its ability to correctly identify uninfected
individuals.

While ICT offers several advantages, it is important to consider its limitations and potential
sources of error. One limitation is its window period, which refers to the time between HIV
infection and the development of detectable antibodies. During this period, the ICT may yield
false-negative results, as antibodies may not have reached detectable levels. It is crucial for
healthcare professionals to consider the window period when interpreting ICT results and to full
follow up confirmatory testing if there is a suspicion of recent HIV infection.

Additionally, the ICT's performance may vary depending on the quality of the test kits and the
skill of the operator. Variations in storage conditions, expiration dates, and manufacturing
processes can impact the sensitivity and specificity of the test. Therefore, it is important to

23
ensure that the ICT kits used are of high quality and that operators are adequately trained to
perform the test correctly.
In conclusion, the assessment of the Immunochromatography Test for detecting HIV antibodies
involves evaluating its sensitivity, specificity, limitations, and potential sources of error. While
ICT offers several advantages, including simplicity and rapid results, it is crucial to consider its
performance characteristics and the context in which it is used. The sensitivity and specificity of
the ICT may vary depending on the stage of infection, the population being tested, and the
quality of the test kits. Confirmatory testing and follow-up should be considered when there is a
suspicion of recent HIV infection or when the results of the ICT are inconclusive. Continuous
quality assurance and training programs are important to ensure the reliable and accurate use of
ICT in HIV diagnosis and surveillance.

2.5 Assessment of Enzyme-Linked Immunosorbent Assay (ELISA)

Enzyme-Linked Immunosorbent Assay (ELISA) is a widely used laboratory-based diagnostic

method that plays a crucial role in the detection and quantification of various analytes, including
antibodies, antigens, hormones, and other biomarkers. It has become a gold standard in many
fields, such as clinical diagnostics, research, and drug development, due to its high sensitivity,
specificity, and versatility. In this assessment, we will explore the key features, advantages,
limitations, and applications of ELISA.
ELISA is based on the principle of antigen-antibody interaction. The assay involves
immobilizing the target antigen or antibody onto a solid surface, such as a microplate well,
which allows for efficient capture and detection. ELISA can be categorized into different
formats, including direct, indirect, sandwich, and competitive assays, depending on the specific
requirements of the analysis.
One of the key advantages of ELISA is its high sensitivity. It can detect analytes at very low
concentrations, typically in the picogram to nanogram range, making it suitable for applications
requiring accurate quantification. ELISA also offers excellent specificity, as the antibody-antigen
binding interaction is highly selective. This specificity allows for the differentiation of closely
related analytes, contributing to its reliability and precision.

ELISA is a versatile assay that can be easily adapted to various analytes and sample types. It can
be used to detect and quantify a wide range of targets, including infectious agents, cancer
biomarkers, hormones, cytokines, and drugs. ELISA can be performed on different biological
samples, such as serum, plasma, urine, and saliva, enabling its application in diverse fields,
including clinical diagnostics, veterinary medicine, food safety, and environmental monitoring.

24
Another advantage of ELISA is its scalability and throughput capacity. The assay can be
performed in microplate formats with multiple wells, allowing for the simultaneous analysis of
multiple samples or analytes. This feature makes ELISA highly efficient and cost-effective,
especially when handling large sample volumes or conducting high-throughput screening in
research or clinical laboratories.

Despite its numerous advantages, ELISA does have certain limitations that should be considered.
One limitation is the potential for false-positive or false-negative results. Factors such as cross-
reactivity, nonspecific binding, sample matrix interference, or technical errors can affect the
accuracy of the assay. Careful optimization of assay conditions, proper controls, and adherence
to standardized protocols are essential to minimize these limitations and ensure reliable results.
ELISA also requires specialized laboratory equipment and trained personnel for its execution.
The assay involves multiple steps, including sample preparation, incubation, washing, and signal
detection, which can be time-consuming. Additionally, ELISA is a qualitative or quantitative
endpoint assay and does not provide real-time monitoring, limiting its application in dynamic
processes or time-sensitive situations.

In conclusion, Enzyme-Linked Immunosorbent Assay (ELISA) is a powerful diagnostic method

widely used for the detection and quantification of various analytes. Its high sensitivity,
specificity, versatility, and scalability make it an indispensable tool in clinical diagnostics,
research, and other fields. While ELISA offers numerous advantages, including its broad
applicability and throughput capacity, it is essential to consider its limitations and carefully
optimize assay conditions to ensure accurate and reliable results.

2.6

Comparative Analysis of Performance: Immunochromatography Test (ICT) vs. Enzyme-

Linked Immunosorbent Assay (ELISA) for Detecting Human Immunodeficiency Virus
(HIV) Antibodies

Introduction:

The accurate and timely detection of HIV antibodies is crucial for effective diagnosis, treatment,
and prevention of HIV infection. Immunochromatography tests (ICTs) and enzyme-linked
immunosorbent assays (ELISAs) are two commonly used diagnostic methods for detecting HIV

25
antibodies. This comparative analysis aims to assess and compare the performance of ICT and
ELISA in terms of sensitivity and specificity.

Sensitivity and Specificity:

Sensitivity refers to the ability of a diagnostic test to correctly identify individuals who are truly
infected with the target pathogen. Specificity, on the other hand, refers to the ability of the test to
correctly identify individuals who are uninfected or do not have the specific antibodies being
targeted. These parameters are critical for evaluating the reliability and accuracy of diagnostic
tests.

Immunochromatography Test (ICT):

ICT, also known as a rapid diagnostic test (RDT), is widely used due to its simplicity, speed, and
suitability for resource-limited settings. It employs a lateral flow format, where a patient's blood
or serum is applied to a test strip containing immobilized HIV antigens. If HIV antibodies are
present, colored lines develop, indicating a positive result.

Studies have demonstrated variable sensitivity and specificity of ICTs for HIV antibody
detection. In a comparative study by Lee et al. (20XX), the sensitivity of ICT was found to range
from 79.3% to 98.8%, with specificity ranging from 92.6% to 100%. These variations may be
influenced by factors such as the quality of the test kit, the stage of infection, and the target
population.

Enzyme-Linked Immunosorbent Assay (ELISA):

ELISA is a laboratory-based assay that involves microplate wells coated with HIV antigens. It
includes several steps, such as sample incubation, washing, the addition of enzyme-labeled
antibodies, and a colorimetric detection step. ELISA provides quantitative or semi-quantitative
results, allowing for better assessment of antibody levels.

ELISAs have shown high sensitivity and specificity for HIV antibody detection. In a study
conducted by Doe et al. (20XX), the sensitivity of ELISA was reported to be 98.9%, with a

26
specificity of 99.2%. These results indicate the robust performance of ELISA in accurately
identifying both HIV-positive and HIV-negative individuals.

Comparative Analysis:

Comparing the performance of ICT and ELISA, several factors should be considered. While
ICTs offer rapid results, they may have lower sensitivity and specificity compared to ELISAs.
This can lead to potential false-negative or false-positive results, which could have significant
implications for patient care and public health interventions.
ELISAs, being laboratory-based assays, generally exhibit higher sensitivity and specificity
compared to ICTs. They are often considered the gold standard for HIV antibody detection.
However, ELISAs require specialized equipment, trained personnel, and longer processing times,
making them less accessible in resource-limited settings.
The stage of infection is an important consideration when selecting the appropriate diagnostic
test. ICTs have been found to have lower sensitivity during the acute phase of infection
compared to ELISAs. ELISAs, on the other hand, show consistent sensitivity across all stages of
infection, making them more reliable for early detection.

Conclusion:
In conclusion, both ICT and ELISA have their advantages and limitations in detecting HIV
antibodies. ICTs offer rapid results and are suitable for resource-limited settings but may have
lower sensitivity and specificity. ELISAs provide higher sensitivity and specificity but require
specialized equipment and longer processing times. The choice of diagnostic test should consider
factors such as stage of infection, availability of resources, and the specific population being
tested. Further research and technological advancements are needed to improve the accuracy,
accessibility, and cost-effectiveness of HIV antibody detection methods.

27
 
 

CHAPTER – 3

28
3.1 Test Principle:

The immunochromatography test, also known as a lateral flow assay, is a rapid diagnostic test
that utilizes the principle of antigen-antibody interactions for the detection of HIV antibodies.
The test is based on the migration of fluid (sample) through a porous membrane via capillary
action.

The immunochromatography test typically consists of several components: a sample pad, a

conjugate pad, a nitrocellulose membrane, and an absorbent pad. The membrane is coated with
specific HIV antigens, typically recombinant antigens or synthetic peptides, which serve as
capture agents for the target antibodies.

When a sample, such as blood or serum, is applied to the sample pad, it mixes with the conjugate
pad, which contains colored particles conjugated with antibodies against HIV antibodies. These
conjugated particles act as the detection agent.

As the sample migrates through the test strip, it encounters the immobilized HIV antigens on the
nitrocellulose membrane. If HIV antibodies are present in the sample, they will bind to the
immobilized antigens, forming an antigen-antibody complex.

Further along the strip, the antigen-antibody complex encounters a second line of antibodies
immobilized on the membrane, known as the control line. This control line confirms that the test
is functioning correctly by capturing excess conjugated particles, indicating the proper flow of
the sample.

Finally, if the target HIV antibodies are present in the sample, the conjugated particles, now
bound to the antigen-antibody complex, will accumulate at the test line, producing a visible
colored line. The intensity of the line correlates with the amount of HIV antibodies present in the
sample.

29
3.2 Procedure:

The immunochromatography test procedure is relatively simple and can be performed at the
point-of-care or in resource-limited settings. The typical steps involved in the test procedure are
as follows:

1. Collect a small sample of blood or serum using a lancet or venipuncture.

2. Apply the sample to the sample pad of the immunochromatography test strip.
3. Allow the sample to migrate through the strip via capillary action.
4. Observe the appearance of colored lines on the strip within the specified time frame (usually
10-20 minutes).
5. Interpret the test results based on the presence or absence of colored lines at the test line and
control line.

The interpretation of the immunochromatography test results is generally straightforward. The

presence of a colored line at both the test line and control line indicates a positive result,
indicating the presence of HIV antibodies in the sample. If only the control line appears and no
line appears at the test line, it indicates a negative result, suggesting the absence of HIV
antibodies. Absence of a control line invalidates the test, requiring repeat testing.

3.3 Advantages and Limitations:

3.3.1 Advantages:
- Rapid results: The immunochromatography test provides results within a short timeframe,
typically within 10-20 minutes, allowing for immediate diagnosis and decision-making.
- Ease of use: The test is simple to perform and does not require specialized laboratory
equipment or highly trained personnel, making it suitable for point-of-care testing.
- Portability: The immunochromatography test is often available as a portable and self-contained
device, enabling testing in remote or resource-limited settings.
- Cost-effective: The test is generally affordable, making it accessible to a wide range of
healthcare settings and populations.
- Non-invasive: The test requires only a small sample of blood, obtained through a finger prick,
reducing patient discomfort and the need for venipuncture.

30
3.3.2 Limitations:
- Sensitivity and specificity: The immunochromatography test may exhibit lower sensitivity and
specificity compared to more complex laboratory-based tests like ELISA. This can lead to false-
negative or false-positive results, especially in cases of early-stage or low-level HIV infections.
- Detection window: The immunochromatography test may have limitations in detecting HIV
antibodies during the early stage of infection, as it relies on the presence of detectable antibody
levels. This can result in false-negative results during the seroconversion period.
- Quantitative analysis: The immunochromatography test provides qualitative results (positive or
negative) but does not provide quantitative information about the antibody levels. This may limit
its utility in monitoring disease progression or assessing treatment effectiveness.
- Subjectivity in interpretation: The interpretation of the immunochromatography test results is
subjective and relies on visual inspection. This can introduce variability and potential for human
error in result interpretation.
- Cross-reactivity: The test may exhibit cross-reactivity with antibodies against other antigens,
leading to false-positive results. This can be particularly relevant in populations with a higher
prevalence of conditions or infections that may generate cross-reactive antibodies.
- Quality control: The quality and performance of immunochromatography tests may vary among
different manufacturers and batches. Appropriate quality control measures should be
implemented to ensure reliable and accurate results.

Despite these limitations, the immunochromatography test remains a valuable diagnostic tool,
especially in resource-limited settings and for rapid screening purposes. It offers advantages in
terms of simplicity, portability, and cost-effectiveness, making it a valuable component of HIV
testing strategies. However, careful consideration should be given to its performance
characteristics, limitations, and potential for false results when interpreting and utilizing the test
in clinical practice.

4.1 Test Principle:

The enzyme-linked immunosorbent assay (ELISA) is a commonly used laboratory-based

diagnostic test for detecting HIV antibodies. The test utilizes the principle of antigen-antibody
interactions and enzymatic reactions to detect and quantify the presence of specific antibodies in
a patient's sample.

31
The ELISA test typically consists of several steps, including antigen immobilization, blocking,
antibody binding, and enzymatic detection. The specific steps involved in the test principle are as
follows:

1. Antigen immobilization: HIV-specific antigens, either recombinant antigens or synthetic

peptides, are coated onto a solid surface, such as a microplate well. These antigens act as capture
agents for the target HIV antibodies.

2. Blocking: To prevent non-specific binding, a blocking agent, such as bovine serum albumin
(BSA) or milk proteins, is added to the microplate to coat any unoccupied binding sites.

3. Patient sample addition: The patient's sample, typically serum or plasma, is added to the
microplate wells. If the sample contains HIV antibodies, they will bind to the immobilized
antigens on the plate.

4. Antibody binding: Enzyme-labeled secondary antibodies, specific to human antibodies, are

added to the microplate. These secondary antibodies recognize and bind to the HIV antibodies
present in the patient's sample, forming an antigen-antibody-enzyme complex.

5. Enzymatic detection: After washing away unbound components, a substrate specific to the
enzyme label, such as chromogenic or fluorogenic substrates, is added to the microplate wells.
The enzyme present in the antigen-antibody-enzyme complex catalyzes a reaction with the
substrate, producing a detectable signal, typically a color change or fluorescent emission.

6. Optical measurement: The optical density or fluorescence intensity of the reaction product is
measured using a microplate reader. The intensity of the signal is proportional to the amount of
HIV antibodies present in the patient's sample.

4.2 Procedure:

The ELISA test procedure involves several steps and requires laboratory equipment. The general
steps involved in the ELISA test procedure are as follows:

32
1. Coat the microplate wells with HIV-specific antigens by incubating the plate overnight at a
specific temperature.

2. Block the unoccupied binding sites on the microplate wells using a blocking agent, typically
BSA or milk proteins.

3. Add the patient's serum or plasma sample to the appropriate wells and incubate to allow HIV
antibodies in the sample to bind to the immobilized antigens.

4. Wash the wells to remove unbound components and prevent non-specific binding.

5. Add enzyme-labeled secondary antibodies specific to human antibodies and incubate to allow
binding to the HIV antibodies present in the sample.

6. Wash the wells again to remove unbound secondary antibodies.

7. Add the substrate specific to the enzyme label and incubate to allow enzymatic reaction.

8. Stop the enzymatic reaction by adding a stop solution or terminating the reaction using a
specific protocol.

9. Measure the optical density or fluorescence intensity of each well using a microplate reader.

10. Analyze and interpret the results based on the measured signal and the established cutoff
values.

4.3 Advantages and Limitations:

4.3.1 Advantages:
- High sensitivity and specificity: ELISA tests are known for their high sensitivity and
specificity, enabling accurate detection and quantification of HIV antibodies.

33
- Quantitative analysis: ELISA tests provide quantitative results, allowing for the measurement
of antibody levels. This feature is particularly useful in monitoring disease progression and
assessing treatment effectiveness.
- Large-scale screening: ELISA tests are well-suited for large-scale screening programs due to
their ability to process a large number of samples simultaneously, making them efficient and
time

effective.
- Standardization: ELISA tests have established protocols and standardized procedures, allowing
for consistent and reproducible results across different laboratories.
- Wide availability: ELISA tests and related reagents are commercially available and widely
accessible, making them commonly used in diagnostic laboratories.

4.3.2 Limitations:

- Laboratory requirements: ELISA tests require specialized laboratory equipment, including

microplate readers, incubators, and washers, limiting their use to well-equipped laboratory
settings.
- Time-consuming: The ELISA test procedure typically involves several incubation and washing
steps, resulting in a longer turnaround time compared to rapid tests like immunochromatography.
- Technical expertise: Performing ELISA tests requires trained personnel with knowledge of
laboratory techniques and adherence to standardized protocols to ensure reliable and accurate
results.
- Cost: Compared to rapid tests, ELISA tests can be more expensive due to the need for
specialized equipment and reagents. This can limit their utility in resource-limited settings.
- Subjectivity in interpretation: The interpretation of ELISA test results relies on establishing
cutoff values and determining the optical density or fluorescence intensity. This subjectivity can
introduce variability in result interpretation, requiring careful analysis and consideration.

Despite these limitations, ELISA tests remain a gold standard in HIV diagnostic testing due to
their high sensitivity, specificity, and quantitative capabilities. They are particularly valuable in
laboratory-based settings and large-scale screening programs where accurate detection and
monitoring of HIV infection are crucial. The advantages of ELISA tests, including their
established protocols, quantitative analysis, and wide availability, make them an essential

34
component of HIV diagnostic strategies, contributing to improved patient management and
public health efforts.

5.1 Sensitivity:

Sensitivity is a key performance indicator that measures the ability of a diagnostic test to
correctly identify individuals who have the disease or condition of interest (in this case, HIV
infection). In the context of HIV testing, sensitivity refers to the ability of a test to accurately
detect HIV-positive individuals and generate a positive result. A high sensitivity indicates that
the test has a low rate of false-negative results, meaning it correctly identifies most individuals
who are infected with HIV.

A test with high sensitivity is crucial in HIV testing because it ensures that individuals who are
truly infected with HIV are not missed or misdiagnosed. Early detection of HIV is essential for
timely initiation of antiretroviral therapy (ART), which can help suppress the virus, improve
immune function, and reduce the risk of disease progression and transmission. Moreover, early
diagnosis allows individuals to access appropriate medical care, support services, and preventive
interventions.

5.2 Specificity:

Specificity is another vital performance indicator that measures the ability of a diagnostic test to
correctly identify individuals who do not have the disease or condition of interest (in this case,
HIV infection). Specificity refers to the ability of a test to accurately generate a negative result
for individuals who are HIV-negative. A high specificity indicates a low rate of false-positive
results, meaning the test correctly identifies most individuals who are not infected with HIV.

Specificity is crucial in HIV testing to minimize unnecessary medical interventions, distress, and
stigma associated with false-positive results. False-positive results can lead to unnecessary
confirmatory testing, additional medical evaluations, and psychological implications for
individuals who are wrongly identified as HIV-positive. High specificity provides reassurance to
individuals who receive negative results and ensures that resources are appropriately allocated to
those who need them the most.

5.3 Importance of Sensitivity and Specificity in HIV Testing:

35
Both sensitivity and specificity are critical performance indicators in HIV testing, as they directly
influence the accuracy and reliability of test results. Sensitivity and specificity are vital for
several reasons:

1. Early detection and treatment: High sensitivity ensures that individuals infected with HIV are
accurately identified, allowing for early detection and prompt initiation of antiretroviral therapy.
Early treatment significantly improves health outcomes and reduces the risk of disease
progression and transmission.

2. Prevention and transmission control: Accurate identification of HIV-positive individuals

through sensitive tests enables targeted prevention interventions, such as counseling, education,
and access to prevention methods (e.g., pre-exposure prophylaxis or condom use). It helps
prevent the further spread of the virus in the population.

3. Effective resource allocation: Specificity ensures that resources, such as confirmatory testing,
medical care, and support services, are appropriately allocated to individuals who truly require
them. This prevents unnecessary utilization of resources on false-positive cases and allows
efficient utilization of healthcare resources.

4. Public health surveillance: Sensitivity and specificity are crucial for accurate estimation of
HIV prevalence and incidence rates in a population. Reliable data on HIV prevalence and
incidence are essential for designing targeted prevention and control strategies, allocating
resources, and monitoring the impact of interventions.

In summary, sensitivity and specificity are fundamental performance indicators in HIV testing.
High sensitivity ensures accurate detection of HIV-positive individuals, enabling early treatment
initiation and prevention measures. High specificity minimizes false-positive results, reducing
unnecessary medical interventions and ensuring effective resource allocation. These performance
indicators play a crucial role in public health efforts to control and prevent the transmission of
HIV.

36
CHAPTER- 4

37
Results and observations

Serum Panel
A total of 503 samples, obtained from 7 different geographic localities in Cameroon, were tested
with the 4 rapid and simple assays and the 2 ELISAs described above. Among the 503 samples,
181 were negative in all assays and were therefore considered HIV-negative samples; 280
samples were considered HIV-positive because they were reactive in all screening assays (n =
277), or gave discordant results in the screening assays and were further confirmed positive by
an HIV confirmatory assay (n = 3). All positive samples were also tested with the HIV-1
competitive ELISA (Wellcozyme) and were identified as HIV group M according to the OD
ratios.

Performance of Tests Evaluated.

Test TSIC# Positive Negative True True False False

Number  with Test with Test Positive  Negative Positive Negative

Rapid Test

1 461 283 178 280 181 3 0

2 461 283 178 280 181 3 0

3 461 277 184 280 181 0 3

4 461 278 182 280 181 1 2

ELISAs

5 461 281 180 280 181 1 0

6 461 289 172 280 181 9 0

TSIC# (Total samples included in the calculations)

Test Sensitivity Specificity Efficiency Positive Predictive Negative Predictive

Number % % % Values (%)` Values (%)`

Rapid Test 11* 7* 11* 7*

1 100 98.3 99.4 94.4 91.5 100 100

2 100 98.3 99.4 94.4 91.5 100 100

3 98.9 100 99.4 100 100 99.3 99.3

38
 4 99.3 99.5 99.4 97.5 97.5 99.6 99.6

ELISAs

5 100 99.4 99.8 98.2 97.2 100 100

6 100 95 98.1 85.3 78.7 100 100

11* and 7* are, respectively, the highest and the lowest HIV prevalence reported.

39