6th International AIDS Conference


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

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A rapid sensitive non-isotopic hybrid-capture assay for HIV nucleic acid sequences.

Int Conf AIDS 1990 Jun 20-23; 6:321 (abstract no. 1029)
Maltzman W, Lee LS, Moore JL, Schochetman G, Ou CY; Enzo Biochem, New York, New York, USA

OBJECTIVE: To develop a sensitive non-isotopic capture system to detect HIV nucleic acids.

METHODS: To capture target nucleic acids, an oligonucleotide sequence derived from a conserved gag or env region of HIV-1 genome was immobilized in a microtiter plate. HIV DNA was captured by the immobilized oligonucleotide via hybridization. Captured DNA was incubated with a second specific signal oligonucleotide; the immobilized complex was detected using an enzyme-dependent amplification system that produced a colorimetric readout.

RESULTS: To test the utility of the hybrid capture assay, we selected peripheral blood mononuclear cells from 85 seropositive and 95 seronegative persons. HIV DNA was amplified by the polymerase chain reaction and was tested using the non-isotopic capture system and the 32P labeled DNA oligomers. The two detection systems gave concordant results. A reconstruction experiment using known amounts of HIV DNA showed the non-isotopic assay system was quantitative.

CONCLUSION: We have developed a rapid, sensitive and non-isotopic capture system to detect HIV-1 nucleic acids. This test can be carried out in a microtiter plate format to facilitate the processing of a large number of specimens in a uniform manner.

Keywords: AEGIS, HIV, HIV-1, Polymerase Chain Reaction, Chimera, Oligonucleotide Probes, HIV Infections, HIV Antibodies, HIV Core Protein p24, HIV-1 Reverse Transcriptase, DNA Probes, Nucleic Acids, HIV Seropositivity, Oligonucleotides, Animal, analysis, ICA6KWDaegis,hiv,hiv-1,polymerasechainreaction,chimera,oligonucleotideprobes,hivinfections,hivantibodies,hivcoreproteinp24,hiv-1reversetranscriptase,dnaprobes,nucleicacids,hivseropositivity,oligonucleotides,animal,analysis,ica6

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1029

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