Document 0827
 DOCN  M9590827
 TI    Analysis and genotyping of PCR products of the Amplicor HIV-1 kit.
 DT    9509
 AU    Barlow KL; Tosswill JH; Clewley JP; Hepatitis and Retrovirus Laboratory,
       Central Public Health; Laboratory, London, UK.
 SO    J Virol Methods. 1995 Mar;52(1-2):65-74. Unique Identifier : AIDSLINE
       GENBANK/Z33480
 AB    The Roche Amplicor PCR kit was used to detect HIV-1 DNA in UK patients
       of known serostatus. Four false-negative and/or equivocal results were
       obtained from patients who were known to be anti HIV seropositive
       (Tosswill et al., 1994). Cells from the blood of these patients were
       shown to contain HIV DNA after extraction, concentration and
       amplification by nested PCR using primers flanking those in the kit. To
       determine whether DNA sequence divergence was the cause of these
       discrepancies, the gag region targeted by the primers in the kit was
       sequenced for specimens giving positive, equivocal and false-negative
       results. No greater degree of sequence divergence was found within the
       primer and probe target regions among the equivocals and false-negatives
       than among the positive control specimens. The few misleading results
       were probably attributable to low copy numbers of proviral DNA in these
       specimens. Sequences obtained from the target and flanking regions of
       the kit were sufficient to allow the genotype of the virus to be
       determined.
 DE    Base Sequence  Comparative Study  DNA Primers  DNA, Viral/*BLOOD
       Electrophoresis, Agar Gel  False Negative Reactions  *Genes, gag
       Genotype  Human  HIV Seropositivity/BLOOD/*DIAGNOSIS
       HIV-1/*GENETICS/*ISOLATION & PURIF  Molecular Sequence Data
       Oligonucleotide Probes  *Phylogeny  Polymerase Chain Reaction/*METHODS
       Reagent Kits, Diagnostic  Sequence Homology, Nucleic Acid  JOURNAL
       ARTICLE

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