Detection of HIV-1-infected cells from patients using nonisotopic in situ hybridization. NLM AIDSLINE Important note: Information in this article was accurate in 1990. The state of the art may have changed since the publication date.

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Detection of HIV-1-infected cells from patients using nonisotopic in situ hybridization.

Blood. 1989 Nov 1;74(6):2295-301. Unique Identifier : AIDSLINE MED/90028766
Singer RH; Byron KS; Lawrence JB; Sullivan JL; Department of Cell Biology, University of Massachusetts Medical; School, Worcester 01655.

Abstract: We have demonstrated that a sensitive, nonisotopic in situ hybridization (ISH) assay can be used to detect HIV-infected cells from seropositive, asymptomatic individuals. Our assay is based on the detection of a biotinated HIV DNA probe hybridized to human immunodeficiency virus (HIV)-infected peripheral blood lymphocytes (PBL) using streptavidin and alkaline phosphatase to identify positive cells. This assay is rapid in that it can be performed within a day and is sensitive enough to unambiguously identify a rare, single, positive cell. Patient samples derived from HIV-seropositive hemophiliacs and HIV-seropositive infants were analyzed before and after coculture with normal PBL. The same samples were investigated using a Dupont P24 antigen-capture kit. It was found that ISH always detected the same positive samples as antigen capture, often in shorter times of coculture. In situ hybridization detected over half of our HIV-infected hemophilia patient population as virus positive, whereas the antigen capture assay detected less than one fourth as virus positive. In situ hybridization detected positive cells directly, without coculture, in 12 out of 35 (34%) hemophiliacs and in three out of eight (37%) infants. The speed, sensitivity, and confidence of ISH and nonisotopic detection indicates that it will be useful as a tool for clinical research and diagnosis.
Keywords: Biotin/DIAGNOSTIC USE Cells, Cultured DNA Probes DNA, Viral/ANALYSIS Human HIV Infections/*DIAGNOSIS HIV Seropositivity/MICROBIOLOGY Lymphocytes/*MICROBIOLOGY Nucleic Acid Hybridization Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. JOURNAL ARTICLEKWDbiotin/diagnosticusecells,cultureddnaprobesdna,viral/analysishumanhivinfections/KWDdiagnosishivseropositivity/microbiologylymphocytes/KWDmicrobiologynucleicacidhybridizationsupport,non-uKWDsKWDgov'tsupport,uKWDsKWDgov't,pKWDhKWDsKWDjournalarticle
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Copyright © 1990 - National Library of Medicine. Reproduced under license with the National Library of Medicine, Bethesda, MD.

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