Human Immunodeficiency Virus

The Virus & The Laboratory Diagnosis

Dr. Sudheer Kher

Prof & Head, Dept of
Microbiology
HIV Genes

Three basic genes

– gag – encodes matrix, capsid, nucleic acid
binding proteins
– pol – encodes reverse transcriptase
– env - encodes surface glycoproteins,
transmembrane proteins
Regulatory genes

 tat ( transactivator of transcription)

 rev (regulator of expression of viral protein)
 nef ( negative expression factor)
 vif (virion infectivity factor)
 vpr (stimulating promoter of virus)
 vpu (HIV-1) & vpx (HIV-2) small viral proteins
promoting maturation and release of progeny
virus from cells.
HIV Antigens

Major surface/envelope glycoproteins

– gp120
– gp41 anchors gp120 to virus
Major capsid proteins
– p24
– P18
Other minor surface & structural proteins
– p55
Laboratory Diagnosis

Test Objective
 Serology
 ELISA Initial Screening
 Latex Agglutination Initial Screening
 Western Blot Confirmatory test
 Immunofluorescence Confirmatory test
 Virion
 RNA, RT-PCR Detection of virus in blood
 P24 antigen Early marker of infection
 Isolation of virus Test not readily available
 Lymphocyte
 CD4:CD8 T cell ratio Correlate of HIV Disease
Laboratory tests for diagnosis of
HIV infection

Screening tests Confirmatory tests

ELISA Western Blot
Rapid tests /Virus isolation
- Latex agglutination /Immunofluorescence
- Dot blot assay Detection of viral nucleic
Simple tests acid
- Particle agglutination - In situ hybridization
- PCR
Detection of p24 antigen
 Before you ask for HIV test
 Are you asking for the test
because you feel it is
mandatory?
 Are you asking for the test
out of fear?
 Have you acquired basic
knowledge of HIV/AIDS?
 Do you value human rights?
Are you aware that your
patient too has rights?
 Are you prepared to do
counseling before asking
for the test?
 Are you prepared to
counsel the patient after
the test?
 Are you prepared to take
informed written consent
of the patient before
sending the sample?
 Are you aware that HIV
test is no substitute for
your precautions?
Strategies for HIV testing in
India
 Strategy-I
 Serum tested by one of
the E/R/S.
 If reactive, sample
considered positive
 If not reactive, sample
considered negative
 Use – Transfusion safety
 Highly sensitive &
reliable kits used
Strategies for HIV testing in
India
 Strategy-II
 Serum reactive with
one E/R/S retested
with second E/R/S
using different Ag/test
principle. If reactive,
reported positive.
 Use – Unlinked
anonymous HIV
Surveillance
Strategies for HIV testing in
India
 Strategy –III
 Serum reactive with two
E/R/S tests is retested
with a third E/R/S.
 Or two of the ERS
followed by confirmatory
test.
 Use – Diagnosis of HIV
infection
-Non-specific tests for HIV

 Blood counts  Diminished CMI –

– Leucopenia Candidial, tuberculin tests.
– Lymphocyte count less  Lab diagnosis of
than 400/cmm
– Thrombocytopenia
opportunistic infections
 T-cell subset assay  Malignancies
– CD4 count below
200/cmm
– Normal CD4:CD8 ratio
2:1. Reversed in AIDS to
0.5:1.
 Hypergammaglobulinaem
ia