6th International AIDS Conference


San Francisco, California, USA — June 20-23, 1990

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Detection of HIV-1-infected cells by nonisotopic in situ hybridization.

Int Conf AIDS 1990 Jun 20-23; 6:338 (abstract no. 1099)
Pingel S, Werchau H, Schmidt E, Richter EO; University of Bochum, Bochum, FRG

OBJECTIVE: A method of nonisotopic in situ hybridization has been developed to detect and localize HIV-1-specific nucleic acid sequences in peripheral blood mononuclear cells (PBMCs) isolated from patients with asymptomatic infection, the AIDS-related complex (ARC) or the acquired immunodeficiency syndrome (AIDS).

METHODS: Experimental conditions were investigated on cytological preparations of in-vitro infected cell culture systems (H9,CEM,PBMCs). DNA probes derived from clone pBH10-RIII were labeled with biotin-dUTP and hybridized to the target sequences. Hybridization sites were subsequently visualized by fluorescence detection methodology. The presence of viral proteins was monitored in parallel by indirect immunofluorescence assays. These methods were employed in a clinical study to determine the proportion of infected cells after isolation.

RESULTS: The sensitivity of the technique was sufficient to detect HIV-1-RNA in cells, HIV-1-DNA within interphase nuclei and few copies of the integrated proviral genome in metaphase spreads. Freshly isolated PBMCs reflected RNA-positive cells at frequencies ranging from 10(-3) to less than 10(-4).

CONCLUSION: The study suggests that in situ hybridization with biotinylated probes may serve as an alternative assay in monitoring the course of infection and in screening clinical specimens for the presence of HIV.

Keywords: AEGIS, HIV-1, Acquired Immunodeficiency Syndrome, HIV Infections, HIV, DNA Probes, HIV Antibodies, HIV Seropositivity, AIDS-Related Complex, Cells, RNA Probes, Biotin, DNA Primers, Leukocytes, Mononuclear, Oligonucleotide Probes, Human, ICA6KWDaegis,hiv-1,acquiredimmunodeficiencysyndrome,hivinfections,hiv,dnaprobes,hivantibodies,hivseropositivity,aids-relatedcomplex,cells,rnaprobes,biotin,dnaprimers,leukocytes,mononuclear,oligonucleotideprobes,human,ica6

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Copyright © 1990 - International AIDS Society (IAS). Reproduction of this abstract (other than one copy for personal reference) must be cleared through the IAS.