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

• Published in: The journal of histochemistry and cytochemistry Vol. 36; no. 12; pp. 1573 - 1577
• Main Authors: Donovan, RM, Cohen, SH, Peterson, WR, Bolton, V, Jordan, GW, Carlson, JR, Vanden Brink, KM, Goldstein, E, Bush, C
• Format: Journal Article
• Language: English
• Published: Los Angeles, CA Histochemical Soc 01.12.1988; SAGE Publications; Histochemical Society
• Subjects: 2-Acetylaminofluorene; Biological and medical sciences; Cell Line; Cytophotometry; DNA Probes; DNA, Viral - analysis; Fundamental and applied biological sciences. Psychology; HIV - genetics; Humans; Immunoenzyme Techniques; Microbiology; Nucleic Acid Hybridization; T-Lymphocytes - microbiology
• ISSN: 0022-1554; 1551-5044
• DOI: 10.1177/36.12.3057074

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.