Malawi Integrated HIV,

Syphilis and Hepatitis B

Rapid Testing

Overview of HIV, Syphilis

and Hepatitis B Rapid
Testing
 Learning Objectives
By the end of this session, participants should be
able to:
 Define Rapid Testing

 Describe sensitivity and specificity in rapid

testing
 Explain advantages of rapid testing

 Define window period

 Explain HIV Nucleic acid amplification testing

 Explain HIV, syphilis and Hepatitis B rapid tests

Definition of Rapid Tests
 Rapid tests are easy-to-use diagnostic tests
that provide quick results, usually in 20
minutes or less.
 Rapid tests directly detect the presence or
absence of antigens or antibody, and are
suitable for point-of care-testing

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 Formats of Rapid Tests

Lateral flow Flow-through

Determine Insti HIV-1/2
Uni-Gold

Control
Control Dot
HIV Test Band HIV Test Dot

Sample pad

Specimen
Flow
HIV, Syphilis and Hepatitis B
Rapid Tests (1)
 HIV, Syphilis and Hepatitis tests detect either
antibodies or virus particles (surface antigen) in
blood.
 Rapid tests have a high degree of accuracy when
administered properly
 More than one test might be needed to confirm
results because of differences in sensitivity and
specificity
HIV, Syphilis and Hepatitis Rapid
Tests (2)
 Some tests are better at detecting positives,
some are better at detecting negatives– this
is called sensitivity and specificity:

 Sensitivity: ability of a test to identify infected

(positive) individuals

 Specificity: ability of a test to identify all un-

infected (negative) individuals
 Antibody/Antigen Rapid Tests

 Detect
i. Antibodies to the antigen (causative agent) which take time
to develop after a person becomes infected (window
period)
ii. Organism’s surface antigens

 Sometimes antibody levels are too low to

register a positive test
 Testing vs Window Period

 Window period is the time between first

infection and when the test can reliably detect
the infection
 The window period length varies across
different tests
 Laboratory tests can detect antibodies against
the causative agent, antigens of the causative
agent, or the actual causative agent.
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Types of Tests used in Malawi

1. Antibody/antigen tests
 Qualitative tests

2. Nucleic Acid Amplification Tests

 Both qualitative and quantitative tests

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Window Period for Antibody
Testing (1)
 Period between infection and detection of HIV
antibody on a lab test
 Window period varies by test and by individual
Time after Percent of infected individuals
infection who will test HIV positive on a
rapid test
4 weeks after 95%
12 weeks after nearly 100%
Window Period for Antibody
Testing (2)
X = HIV
O = HIV Antibody
X X
X X
O O
X X O X
X X X
X X X
X X
O
X X
O X
X X
X X
O
X X
X
O X
X O

Time: Time:
Initial HIV 4-12 weeks after
infection infection
Window Period for Antibody
Testing (3)
 The window period for syphilis is between 3-6 weeks
 Window period for Hepatitis B surface antigens is
(HBsAg) 1-4 weeks
 Nucleic Acid Amplification
Tests
 Detect parts of the actual virus
 Can be effective 2 to 3 weeks after infection
 Detect parts of the virus before antibodies can be
detected on an Rapid Test
 DNA PCR tests
 Useful for diagnosing infants under 12 months old
 Dried blood spot collection is the most common
method for testing

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Types of HIV Antibody Tests
Used in Malawi

 Types of antibody tests used:

 Determine
 UniGold
 SD Bioline
 Oraquick (HIV Self testing)
 Asante (HIV Recency testing)
 All are rapid tests– results are ready in 20
minutes or less
 Types of HIV Antibody Test
Devices

Determine UniGold Asante OraQuick SD Bioline HIV

Types of Hepatitis Antibody Test
Devices

SD Bioline HBsAg
DetermineTM HBsAg
Types of Syphilis Antibody
Test Devices

DetermineTM Syphilis TP
SD Bioline Syphilis

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How Oral fluid based Tests Work
 Oral fluid based test uses oral fluid to test for the
presence of HIV antibodies
 The oral fluid based test are used for self testing
 An individual collects oral fluid using the test
device, performs a test and interprets the test result
themselves
 When antibodies to HIV are present, a positive
reaction is indicated by a pink/redline on both
test and control window.
 Testing for Syphilis
 Syphilis rapid test is a qualitative test used
to detect antibodies against Treponema
pallidum found in blood
 Tests have a high degree of accuracy when
administered properly
 SD Bioline Syphilis 3.0 is the current rapid
test used in Malawi.

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SD Bioline Syphilis 3.0 Kit
How Syphilis blood based
Rapid Tests Work
 Dispense 20µL of blood on a test device,
then add assay diluent for Bioline Syphilis
or Chase Buffer for DetermineTM Syphilis as
recommended.
 Blood combines with buffer then moves
laterally along the test strip
 When antibodies to Treponema pallidum
are present, a positive reaction is indicated
by a pink/red line on both test and control
window.
 Testing for Hepatitis B Virus
 Hepatitis B rapid test is a qualitative test
used to detect Hepatitis B surface antigen
in blood.
 Tests have a high degree of accuracy
when administered properly.
 Determine HBsAg 2 is the current rapid test
used in Malawi.

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Hepatitis B rapid Test Kit

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How Hepatitis B Virus blood based
Rapid Test Works - Determine HBsAg
 Dispense 50 μL of sample (precision pipette)
to the middle of the sample pad (marked by
the arrow symbol).
 Apply one drop of Chase Buffer to the sample
pad
 When the surface antigens of Hepatitis B virus
are present, a positive reaction is indicated by a
red control line on control window and a black
line on the test window
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 Key Points
 Antibody/antigen tests detect
antibodies/organism’s surface antigens
 Nucleic acid amplification tests detect parts of
the actual organism
 Antibody tests used in Malawi:
 Determine, UniGold, SD Bioline, Oraquick, and
Asante, Syphilis and Hep B
 Syphilis rapid test detects antibodies against
Treponema pallidum
 Hepatitis B rapid test detects the Surface
antigen of Hepatitis B Virus