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M94A2703.TXT
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1994-10-25
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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
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).