CHAPTER 2: HIV TESTING TECHNOLOGIES

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 Enabling objectives
At the end of this section participants will be able to.

• Explain the Spectrum of HIV testing Technologies

• Identify complexity of HIV Testing technologies

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 Outlines:
 Spectrum of HIV Testing Technologies

 Complexity of HIV Tests

 Chapter Summary

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 Spectrum of HIV testing Technologies
• .

Activity 2.1. Self-Reflection

Instruction: Reflect your ideas on the following
questions

List the current HIV testing technologies

Time: 5 minutes

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 Spectrum cont.….
 The period immediately following HIV infection is the eclipse period.

 During eclipse period no assay can detect HIV as;

The amount of nucleic acid from the virus is miniscule

 Low amount of antibodies are produced only in response to the

presence of the virus

 The eclipse period typically lasts approximately 10 days.

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 Spectrum cont.….
 HIV testing technologies categorized as serological and Virological

assays.

 Virological assays can detect as early as 12 days of HIV infection.

 Whereas serological assays need nearly 18 days to detect the

concentrations of antibodies in the body fluid.

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 Spectrum cont.….
.

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 Spectrum cont.….
 The length of HIV detection period affected by
 The type of serological assay used,
An individual’s immune response,
The type of the body fluid used.

 Based on the composition of the antigen and what the assay detects
serological assays categorized in to four generations .

 Fourth generation assays that detect both HIV p24 Ag and Ab can
potentially identify HIV-infected individuals during acute infection
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 Spectrum cont.….
.

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 Spectrum cont.….
• Serological Assays
Activity 2.2. Think/Pair/share
Instruction: First take a while to think on the
following questions and be in pair with a partner
seating next to you and share your idea regarding
the question and reflect what you have discussed
to the larger group.
1.1.1. What are the three Phenomenon’s on the
serological reaction?
Time: 4 min to think and share to your partner and
1 minute for reflection

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 Serological Assays
 There are three Phenomenon's on Antigen-antibody reactions

Pro zone phenomenon- false negative resulting from excess antibody

titer which interferes with formation of Antigen-antibody lattice

Post zone phenomenon- is false negative resulting from excess

antigen titer

Equivalence zone – proportional amount of antibody and Antigen

(optimal ratio of Antigen to Antibody)

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 Rapid Diagnostic Test:
 HIV rapid tests are qualitative assays that detect HIV antibodies.

 Most of them can detect HIV-1 and HIV-2 and they are as reliable
as EIAs.

 There are three main formats or types of rapid HIV tests

 Immuno-concentration (flow-through device

 Immuno-chromatography (lateral flow)

 Particle agglutination

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 Immuno-concentration (flow-through device)

 Immuno-concentration devices
are usually cartridges with HIV
antigen attached to a membrane.

 The specimen and individual

reagents are each added to the
cartridge in a series of steps.

 Presence of HIV Ab is indicated

by colored spot or line.
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 Immuno- chromatography
 Specimen is applied to a pad (filter)
where it mixes with gold or selenium
Add
colloid- antigen conjugate. Sample
Test
Control
Conjugate

This mix migrates through the

Line
 Line

nitrocellulose strip to immobilized

recombinant antigens and synthetic
peptides at the patient window.

 If HIV Ab are present, a red line will

form in the test area of the strip
.
Eg One step anti-HIV (1&2), First response
HIV-1, Uni-Gold HIV, Stat pack

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 Particle agglutination
 Agglutination assays were among
the first of the rapid tests developed.

 The round circles represent antigen-

coated latex particles that bind to
antibodies to HIV (represented by
the “Y”).

 Agglutination or clumping occurs

when the antibodies bind to the
antigen-coated particles. Eg, Capilus
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 HIV Rapid Tests, Advantages
 Increase access to prevention
 Test time under 30 minutes
 Support increased number of
 Most require no refrigeration
testing sites

 Encourage self-testing  None or one reagent

 Same-day diagnosis and  Minimal or no equipment required
counseling  Minimum technical skill
 Robust and easy to use
 Ability to use whole blood

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 HIV rapid tests disadvantages
 Small numbers for each test run

 Quality Assurance/Quality Control at multiple sites

 Test performance varies by product

 Reader variability in interpretation of results

 Limited end point stability of the results

 Need refrigerator to store the kits Eg Capilus

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 Enzyme immunoassays (EIAs)
 EIA is a quantitative assay that measure HIV Ab.
 Most EIAs can detect antibodies to HIV-1 and HIV-2.
 Here is how EIA works:

Sample is added to micro well plate that has been

coated with HIV antigen(s)
After a series of reagent additions, incubations
and washings, the plate is placed in reading
device.

The reading device measures the optical density of color that developed

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 Virological assays
 HIV DNA PCR is the most widely used initial assay for early infant diagnosis.

 HIV RNA PCR and other nucleic acid detection techniques assist in clinical
management.

 P24 Core protein- Used for the diagnosis of pediatric HIV-1 infections and
blood bank safety (high incidence countries)
Can be detected,
 2 to 3 weeks after HIV infection
About 6 days before antibody tests become reactive
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 Complexity of HIV Tests
• .

Activity 2.2. Self-Reflection

Instruction: Reflect your ideas on the following
question.

What are the basic criteria to categorize the

complexity of HIV tests

Time: 3 minutes

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 Complexity of HIV Tests
 Varies, level 1 to level 4, in terms of equipment
and technical skill.

Level 1: No additional equipment and little or no

laboratory experience needed

Level 2: Reagent preparation or a multi-step

process is required; centrifugation or optimal
equipment

Level 3: Specific skills such as diluting are required

Level 4: Equipment and trained laboratory technician are required
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 Challenges of HIV Testing
 The ability of some test to detect early infections is suboptimal.

 Specialized testing is required to diagnose HIV infection in infants younger than

18 months.

 Some tests may not be able to detect antibodies produced against specific HIV
subtypes.

 Cross reactivity with other health conditions or infections

 Some technologies require specific equipment that must be properly maintained.

 Personnel need a certain level of skill to accurately perform and interpret tests
varies (from minimal to high level
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 Chapter Summery
• Spectrum of HIV Diagnostic categorized as serological and
Virological assays

• Serological HIV testing technologies characterized as rapid and EIA.

• There are three formats of HIV Rapid tests

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 References:
1) Consolidated guidelines on HIV prevention, testing, treatment, service delivery and
monitoring: recommendations for a public health approach. Geneva: World Health
Organization; 2021. Licence: CC BY-NC-SA 3.0 IGO

2) Consolidated guidelines on HIV testing services, 2019. Geneva: World Health

Organization; 2020. Licence: CC BY-NC-SA 3.0 IGO.

3) Web Annex I. In vitro diagnostics for HIV diagnosis. In: Consolidated guidelines on
HIV testing services, 2019. Geneva: World Health Organization; 2020. Licence: CC
BY-NC-SA 3.0 IGO.

4) WHO, Guidance note on the selection of technology for the early diagnosis of HIV in
infants and children
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.

QUESTIONS????

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