PubMed ID

2804364

UMMS Affiliation

Department of Cell Biology; Department of Pediatrics

Date

11-1-1989

Document Type

Article

Subjects

Biotin; Cells, Cultured; DNA Probes; DNA, Viral; HIV Infections; HIV Seropositivity; Humans; Lymphocytes; Nucleic Acid Hybridization

Disciplines

Cell Biology | Life Sciences | Medicine and Health Sciences

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.

Rights and Permissions

Citation: Blood. 1989 Nov 1;74(6):2295-301. Link to article on publisher's website

Related Resources

Link to article in PubMed