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

Robert H. Singer, K. S. Byron, J. B. Lawrence, J. L. Sullivan

Research output: Contribution to journalArticle

18 Citations (Scopus)

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 homophiliacs 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.

Original languageEnglish (US)
Pages (from-to)2295-2301
Number of pages7
JournalBlood
Volume74
Issue number6
StatePublished - 1989
Externally publishedYes

Fingerprint

Viruses
In Situ Hybridization
HIV-1
HIV
Coculture Techniques
Assays
Antigens
Lymphocytes
Cells
Blood
Streptavidin
DNA Probes
Hemophilia A
Alkaline Phosphatase
Research
Population

ASJC Scopus subject areas

  • Hematology

Cite this

Singer, R. H., Byron, K. S., Lawrence, J. B., & Sullivan, J. L. (1989). Detection of HIV-1-infected cells from patients using nonisotopic in situ hybridization. Blood, 74(6), 2295-2301.

Detection of HIV-1-infected cells from patients using nonisotopic in situ hybridization. / Singer, Robert H.; Byron, K. S.; Lawrence, J. B.; Sullivan, J. L.

In: Blood, Vol. 74, No. 6, 1989, p. 2295-2301.

Research output: Contribution to journalArticle

Singer, RH, Byron, KS, Lawrence, JB & Sullivan, JL 1989, 'Detection of HIV-1-infected cells from patients using nonisotopic in situ hybridization', Blood, vol. 74, no. 6, pp. 2295-2301.
Singer, Robert H. ; Byron, K. S. ; Lawrence, J. B. ; Sullivan, J. L. / Detection of HIV-1-infected cells from patients using nonisotopic in situ hybridization. In: Blood. 1989 ; Vol. 74, No. 6. pp. 2295-2301.
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