Document 2703
 DOCN  M94A2703
 TI    An approach to monitor viable HIV-1: (II). Non-isotopic microtiter plate
       reverse transcriptase assay for drug susceptibility of clinical HIV
       isolates.
 DT    9412
 AU    Saito T; Suzuki K; Takahashi T; Kano K; Kondo M; Hayashi T; Imai M;
       Kanagawa Prefectural Institute of Health, Yokohama, Japan.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(1):244 (abstract no. PB0405). Unique
       Identifier : AIDSLINE ICA10/94369872
 AB    OBJECTIVES: We applied non-isotopic reverse transcriptase assay (RT
       assay) for quantitative analysis of HIV cultured with PBMC to study
       susceptibility of HIV isolates to antivirals. RT assay was compared with
       p24 antigen assay and with the observation of cytopathic effect (CPE).
       METHODS: Two strains of HIV-1, AZT resistant and sensitive, were
       cultured using AZT and ddI with HIV negative PBMC for a week. The
       samples were taken out at 1, 3, 5 and 7 days after the infection for the
       analysis of RT, p24, and CPE tests. For the RT assay 20 microliters of
       cultured supernatant was directly used for its measurement. RESULTS: AZT
       sensitive HIV showed low RT activity in the presence of AZT or ddI. AZT
       resistant HIV showed the higher RT activity as the days went on after
       infection in the present of AZT but its activity was kept at low value
       in the presence of ddI. CPE observation corresponded with these RT
       activities. DISCUSSION AND CONCLUSION: This non-isotopic RT assay was
       simple and quantitative analysis became possible. One plate was able to
       handle 96 samples at one time. The RT assay was proved to be valuable
       method to evaluate drug susceptibility of HIV.
 DE    Antiviral Agents/*PHARMACOLOGY  Cytopathogenic Effect, Viral/DRUG
       EFFECTS  Didanosine/PHARMACOLOGY  Human  HIV Core Protein p24/ANALYSIS
       HIV-1/*DRUG EFFECTS  Reverse Transcriptase/ANALYSIS  Virus Cultivation
       Zidovudine/PHARMACOLOGY  MEETING ABSTRACT

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