AEGiS-06IAC: Detection of HIV-1 DNA by PCR using structural and regulatory genes on blood samples from seronegative high risk patients.

6th International AIDS Conference

San Francisco, California, USA — June 20-23, 1990

Detection of HIV-1 DNA by PCR using structural and regulatory genes on blood samples from seronegative high risk patients.

Int Conf AIDS 1990 Jun 20-23; 6:339 (abstract no. 1100)
Dawood MR, Allan R, Stackiw W, Conway B, Bechtel LJ, Hirsch MS, Hammond G; Cadham Provincial Laboratory, Winnipeg, MB, Canada

OBJECTIVES: To study the use of oligonucleotide primers from gag, pol, and vif genes for the detection of HIV-1 DNA in blood samples from seronegative high risk patients.

METHODS: DNA was extracted from PBLs collected from 10 seronegative staff without risk factor for HIV-1 infections, 38 high risk seronegative patients and 22 seropositive patients. One microgram aliquots from each patient were used for the detection of HIV-1 DNA using the PCR technique. Oligonucleotide primers from the conserved regions of the gag, pol, and vif genes were used for PCR. After 30 cycles of PCR using Taq polymerase, DNA was southern-blotted. Membranes were hybridized with 32P probes, then autoradiographed. DNA specimens under code from high risk patients were tested in parallel with PCR at M.G.H. using another set of primers from the pol gene and an alkaline phosphatase linked probe. A sample was considered positive with at least 3 positive primers out of 4, (-) if negative with at least 3 primers and (+/-) if positive with only 2 out of 4 primers.

RESULTS: TABULAR DATA, SEE ABSTRACT VOLUME.

CONCLUSIONS: It is important to verify HIV-1 PCR results with more than one set of primers and probes. The criteria for interpretation of PCR results require further standardization.

Keywords: AEGIS, Polymerase Chain Reaction, HIV-1, DNA Primers, HIV Infections, Genes, pol, Risk Factors, Taq Polymerase, Genes, vif, Genes, Regulator, Human, genetics, ICA6

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