       Document 0006
 DOCN  M9460006
 TI    Recombinant virus assay: a rapid, phenotypic assay for assessment of
       drug susceptibility of human immunodeficiency virus type 1 isolates.
 DT    9404
 AU    Kellam P; Larder BA; Antiviral Therapeutic Research Unit, Wellcome
       Research; Laboratories, Beckenham, Kent, United Kingdom.
 SO    Antimicrob Agents Chemother. 1994 Jan;38(1):23-30. Unique Identifier :
       AIDSLINE MED/94190047
 AB    Antiviral drug susceptibility assays for clinical human immunodeficiency
       virus type 1 (HIV-1) isolates are required to monitor the development of
       drug resistance during clinical trials and antiretroviral drug therapy.
       First-generation phenotypic assays possess a number of drawbacks, not
       least the selection of unrepresentative virus populations during
       cocultivation. Here we describe a rapid phenotypic assay for the
       assessment of the susceptibility of clinical isolates to reverse
       transcriptase (RT) inhibitors. This procedure, called the recombinant
       virus assay, allows the generation of viable virus by homologous
       recombination of a PCR-derived pool of RT coding sequences into an
       RT-deleted, noninfectious proviral clone, pHIV delta BstEII. A nested
       PCR procedure has been optimized to allow the amplification of an RT
       pool from both uncultured and cocultured infected patient peripheral
       blood lymphocyte (PBL) DNA for subsequent use in the creation of
       recombinant viruses. Analysis of two patients during the course of
       zidovudine therapy showed that this approach produced viruses which
       accurately exhibited the same genotype and phenotype as that of the
       original infected PBL DNA. The recombinant virus assay can be performed
       in approximately 3 weeks without the use of donor PBLs and therefore
       represents a rapid, nonselective procedure for the assay of clinical
       isolates.
 DE    Antigens, CD4/ANALYSIS  Antiviral Agents/*PHARMACOLOGY  Base Sequence
       Cell Line  DNA, Viral/BIOSYNTHESIS  Genotype  Hela Cells  Human  HIV
       Infections/*MICROBIOLOGY  HIV-1/*DRUG EFFECTS/GENETICS
       Lymphocytes/METABOLISM/MICROBIOLOGY  Microbial Sensitivity
       Tests/*METHODS  Molecular Sequence Data  Phenotype  Plaque Assay
       Polymerase Chain Reaction  Recombination, Genetic  Virus
       Replication/DRUG EFFECTS  Zidovudine/PHARMACOLOGY  JOURNAL ARTICLE

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

