OBJECTIVETo establish a rapid assay for assessment of drug susceptibility of human immunodeficiency virus type 1 isolates (Recombinant virus assay).

METHODSThis procedure allows the generation of viable virus with SI phenotype by homologous recombination of a RT-PCR-derived pool of reverse transcriptase (RT) coding sequences into an RT-deleted, noninfectious proviral clone, pHIV delta RTBstE II. Then the drug susceptibility of recombinant virus to RT inhibitors can be assessed in the Hela CD4+ plaque reduction assays.

RESULTSAnalysis of 7 HIV strains with SI or NSI phenotype showed that recombinant viruses accurately exhibited the same genotype as that of the original HIV1 isolates. The results of drug susceptibilities of HIV1 isolate got by recombinant virus assay were the same as that by standardized peripheral blood mononuclear cell culture assay.

CONCLUSIONRecombinant virus assay is a rapid and accurate method to assess the drug sensitivity of HIV1 isolates with SI or NSI phenotype.

Antiviral Agents ; pharmacology ; CD4 Antigens ; analysis ; DNA, Viral ; biosynthesis ; HIV Infections ; virology ; HIV-1 ; drug effects ; genetics ; HeLa Cells ; Humans ; Microbial Sensitivity Tests ; methods ; Phenotype ; Recombination, Genetic ; Virus Replication ; drug effects