In situ detection of human immunodeficiency virus (HIV) nucleic acid in H9 cells using nonradioactive DNA probes and an image cytophotometry system

R. M. Donovan, Stuart H Cohen, W. R. Peterson, V. Bolton, G. W. Jordan, J. R. Carlson, K. M. Vanden Brink, E. Goldstein, C. Bush

Research output: Contribution to journalArticle

Abstract

Rapid and sensitive nonradioactive methods to detect human immunodeficiency virus (HIV)-infected cells are needed in clinical medicine. We developed an in situ hybridization test using 2-acetylaminofluorene (AAF)-labeled HIV DNA as a hybridization probe. Hybridized probe was detected using rabbit anti-AAF antibody, followed by alkaline phosphatase-conjugated goat anti-rabbit, and the bromochloroindolyl phosphate-nitroblue tetrazolium reaction. An image cytophotometry system was used to quantitate the percentage of HIV-infected cells. These methods were used to determine the percentage of H9 cells infected with HIV. HIV was detected in 0% of cells on day 1 post infection, 7% on day 4, 41% on day 8, and 5% on day 15. These results paralleled those of the reverse transcriptase assay and an antigen capture ELISA assay for HIV antigen. Thus the AAF modified HIV DNA probe detected HIV nucleic acid in infected H9 cells and the image cytophotometry system improved the sensitivity and objectivity of detection.

Original languageEnglish (US)
Pages (from-to)1573-1577
Number of pages5
JournalJournal of Histochemistry and Cytochemistry
Volume36
Issue number12
StatePublished - 1988

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Cytophotometry
DNA Probes
Nucleic Acids
HIV
2-Acetylaminofluorene
Rabbits
Nitroblue Tetrazolium
Antigens
Clinical Medicine
RNA-Directed DNA Polymerase
Goats
In Situ Hybridization
Alkaline Phosphatase
Enzyme-Linked Immunosorbent Assay
Phosphates
Antibodies
DNA

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

In situ detection of human immunodeficiency virus (HIV) nucleic acid in H9 cells using nonradioactive DNA probes and an image cytophotometry system. / Donovan, R. M.; Cohen, Stuart H; Peterson, W. R.; Bolton, V.; Jordan, G. W.; Carlson, J. R.; Vanden Brink, K. M.; Goldstein, E.; Bush, C.

In: Journal of Histochemistry and Cytochemistry, Vol. 36, No. 12, 1988, p. 1573-1577.

Research output: Contribution to journalArticle

Donovan, RM, Cohen, SH, Peterson, WR, Bolton, V, Jordan, GW, Carlson, JR, Vanden Brink, KM, Goldstein, E & Bush, C 1988, 'In situ detection of human immunodeficiency virus (HIV) nucleic acid in H9 cells using nonradioactive DNA probes and an image cytophotometry system', Journal of Histochemistry and Cytochemistry, vol. 36, no. 12, pp. 1573-1577.
Donovan, R. M. ; Cohen, Stuart H ; Peterson, W. R. ; Bolton, V. ; Jordan, G. W. ; Carlson, J. R. ; Vanden Brink, K. M. ; Goldstein, E. ; Bush, C. / In situ detection of human immunodeficiency virus (HIV) nucleic acid in H9 cells using nonradioactive DNA probes and an image cytophotometry system. In: Journal of Histochemistry and Cytochemistry. 1988 ; Vol. 36, No. 12. pp. 1573-1577.
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