A rapid method for detection and differentiation of amplified HIV-1 and HIV-2 DNA using a sensitive chemiluminescent DNA probe system (HPA).

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A rapid method for detection and differentiation of amplified HIV-1 and HIV-2 DNA using a sensitive chemiluminescent DNA probe system (HPA).

Int Conf AIDS. 1990 Jun 20-23;6(1):136 (abstract no. Th.A.10). Unique Identifier : AIDSLINE ICA6/10001090
McDonough S; Brown V; Ryder T; Ou CY; Schochetman G; Gen-Probe, Inc., San Diego, California, USA

Abstract: OBJECTIVE: As the occurrence of HIV-2 infection increases among the population, the need for differentiation of HIV-1 and HIV-2 becomes more important. The objective of this work was to apply the Hybridization Protection Assay (HPA) to the differentiation of amplified HIV-1 and HIV-2 DNA. METHODS: HPA using DNA probes directly labeled with acridinium ester, and a simple homogeneous assay format, was applied to the detection of amplified HIV-1 and HIV-2 DNA. HIV-1 and HIV-2 DNA was amplified with 35 cycles of standard PCR using HIV-1 gag-region primers SK38/SK39 or primers representing conserved regions in the vpx gene of HIV-2. RESULTS: DNA from 25 or more HIV-1 infected cells amplified with HIV-1 primers gave signal significantly above background when hybridized with the HIV-1 DNA probe mix (greater than or equal to 95x). When two amounts of lysate (differing 10 fold) from HIV-2 infected cells were amplified with this primer pair, a low level signal was seen (6x background). HIV-2 DNA in lysate from infected cells was easily detected above background following amplification with HIV-2 primers and hybridization to the HIV-2 DNA probe mix. No cross reactivity was seen when the HIV-2 probe mix was hybridized to DNA from 8000 HIV-1 infected cells amplified with the HIV-2 primer pair. CONCLUSIONS: The amplification of HIV-1 and HIV-2 sequences, followed by detection in the HPA format with the chemiluminescent DNA probes described here, provides a rapid and sensitive means for detection and differentiation of these viruses.

Keywords: *DNA Probes DNA, Viral/*ANALYSIS Gene Amplification Human HIV-1/*GENETICS/ISOLATION & PURIF HIV-2/*GENETICS/ISOLATION & PURIF Nucleic Acid Hybridization Polymerase Chain Reaction ABSTRACT
901230
M90C3708

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