Comparison of non-radioactive DNA hybridization probes to detect human immunodeficiency virus nucleic acid. NLM AIDSLINE Important note: Information in this article was accurate in 1988. The state of the art may have changed since the publication date.

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Comparison of non-radioactive DNA hybridization probes to detect human immunodeficiency virus nucleic acid.

Mol Cell Probes. 1987 Dec;1(4):359-66. Unique Identifier : AIDSLINE MED/88275999
Donovan RM; Bush CE; Peterson WR; Parker LH; Cohen SH; Jordan GW; Vanden Brink KM; Goldstein E; Department of Internal Medicine, University of California, Davis; 95616.

Abstract: Simple and sensitive methods to directly detect the human immunodeficiency virus (HIV) are needed for routine use in the clinical laboratory. In this study, we compared DNA probes prepared by: (1) nick translation with biotinylated dATP; (2) direct covalent biotinylation with photobiotin; (3) direct covalent reaction with 2-acetylaminofluorene (AAF); and (4) a standard radioactive (32P) nick translation procedure. These four DNA probes were hybridized with dilutions of purified target HIV DNA blotted onto nitrocellulose strips. Hybridization was detected using a complex of strepavidin-alkaline phosphatase [for (1) and (2)], alkaline phosphatase-tagged antibodies [for (3)] and by autoradiography [for (4)]. Alkaline phosphatase was detected colorimetrically using nitroblue tetrazolium and 5-bromo-4-chloro-3-indolyl phosphate. After 1 h, AAF probes were most sensitive (amount detected less than 5 pg), followed by biotin (10 pg), photobiotinylated probes (20 pg) and the radioactive probe (10 pg). The AAF probes were then used to detect HIV DNA in infected CEM cells. We conclude that non-radioactive DNA labelling methods can be used to directly detect HIV DNA under conditions compatible with present clinical laboratory procedures.
Keywords: Biotin/DIAGNOSTIC USE Comparative Study DNA, Viral/*ANALYSIS Human HIV/*GENETICS/ISOLATION & PURIF *Nucleic Acid Hybridization Phosphorus Radioisotopes Support, U.S. Gov't, P.H.S. 2-Acetylaminofluorene/DIAGNOSTIC USE JOURNAL ARTICLE

KWDbiotin/diagnosticusecomparativestudydna,viral/KWDanalysishumanhiv/KWDgenetics/isolation&purifKWDnucleicacidhybridizationphosphorusradioisotopessupport,uKWDsKWDgov't,pKWDhKWDsKWD2-acetylaminofluorene/diagnosticusejournalarticle
881030
M88A0626

Copyright © 1988 - National Library of Medicine. Reproduced under license with the National Library of Medicine, Bethesda, MD.

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