HIV-1 detection by PCR and alkaline phosphatase directly labeled DNA probes.

Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.

HIV-1 detection by PCR and alkaline phosphatase directly labeled DNA probes.

Abstr Gen Meet Am Soc Microbiol. 1993;93:434 (abstract no. T-12). Unique Identifier : AIDSLINE ASM93/93291815
Castro M; Bishr M; Tang K; Vela R; University of North Texas, Denton.

Abstract: A DNA amplification procedure using a heat stable DNA Polymerase and the polymerase chain reaction is described for the detection of HIV-1. A set of primers was designed to obtain a 500 b.p. amplification product from the gag region of a plasmid containing an inactivated version of HIV-1. Using this set of primers in conjunction with the polymerase chain reaction, a 500 b.p. product is obtained. Following immobilization by dot-blotting on a nylon membrane, the HIV-1 samples were hybridized with a DNA probe that was covalently coupled to Alkaline Phosphatase. Upon exposure of the membrane to dioxetane chemiluminescent substrate, we can detect less than 10(3) HIV-1 targets within 60 minutes. These results suggest that amplification of specific sequences within the HIV-1 gene by the polymerase chain reaction, coupled with non-radioactive hybridization detection, may provide a highly sensitive and specific method for the detection of HIV-1 in the early stages of viral infection, where immunological methods fail to detect the HIV-1 virus.

Keywords: Acquired Immunodeficiency Syndrome/*DIAGNOSIS/MICROBIOLOGY Alkaline Phosphatase Chemiluminescence *DNA Probes Genes, gag Human HIV-1/GENETICS/*ISOLATION & PURIF Plasmids Polymerase Chain Reaction/*METHODS ABSTRACT
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M9391156

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