lab diagnosis of hiv · 2 the role of proviral dna tests • hiv-1 proviral dna can be detected by...

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Dr Sarah Sasson Immunopathology Registrar

HIV, Immunology and Infectious Diseases Department and SydPath, St Vincent's Hospital

HIV: Basic Virology

Courtesy of Philip Cunningham

HIV testing @ SydPath

1)  Architect 4th generation chemiluminescence assay

•  Detects p24 Ag and Ab to HIV-1 and HIV-2

2)  (if +) Genscreen

•  Detects Ab to HIV-1 and HIV-2

3)  (if +) p24 ELISA & HIV Western Blot

Courtesy Philip Cunningham, St Vincent's Hospital Sydney

Evolution of HIV Western Blot

•  Highly Specific

•  Less Sensitive

•  Expensive $$$

•  Results may be:

  POSITIVE

  NEGATIVE

  INDETERMINATE

DNA PCR RNA PCR

p24 Ag

1wk 2wk 3wk 2mo 6mo 1yr 2yr 3yr +8yr

early recent/established late

Evolution: antibody/ antigen testing response

Courtesy of Philip Cunningham

The role of proviral DNA tests

•  Detection of HIV-1 proviral DNA by PCR can provide early evidence of HIV-1

infection (approximately 10-14 days after infection), when results of routine

diagnostic assays are inconclusive.

•  The linear cDNA transcripted prior to integration is the “proviral” DNA.

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The role of proviral DNA tests

•  HIV-1 proviral DNA can be detected by a PCR that targets a segment of the highly

conserved HIV-1 gag gene or LTR region. Clinical studies have indicated this is

highly sensitive (>95%) and specific (>98%).

•  Currently SydPATH is the only site licensed in Australia for this test

•  The limit of detection is 10 copies/μL

•  Research assays can detect 0.3 copies/ μL

HIV Monitoring and management tests

•  CD4 count and percentage

•  HIV VL

•  Viral genotype

•  Viral resistance

•  Viral tropism

•  HLA B57