Document 0826
 DOCN  M9590826
 TI    The detection of HIV-1 proviral DNA by PCR in clotted blood specimens.
 DT    9509
 AU    Boriskin YuS; Booth JC; Fernando S; Carrington D; Evans MR; Hay P;
       Coates AR; Department of Medical Microbiology, St George's Hospital
       Medical; School, London, UK.
 SO    J Virol Methods. 1995 Mar;52(1-2):87-94. Unique Identifier : AIDSLINE
       MED/95286752
 AB    Six hundred and ninety-two specimens, each consisting of a suspension of
       the residual free cells in samples of clotted blood (but not the clots
       themselves) from 680 patients at high risk for exposure to human
       immunodeficiency virus (HIV), were tested by a nested polymerase chain
       reaction (PCR) procedure which had been optimized to give a sensitivity
       of detection of one copy of HIV-1 proviral plasmid DNA, and the results
       were compared to those of testing for antibody to HIV on the same
       specimens. Fifty-one of the specimens were positive for antibody to HIV
       and 49 of these were also positive by the PCR; the two samples which
       gave discordant results were found to be PCR-positive when the test was
       repeated on DNA extracted from the clots themselves. Two specimens were
       found to be negative for antibody to HIV but were positive by the PCR
       (53 positive specimens in all). Direct sequencing of the PCR DNA
       products confirmed their specificity in all cases and demonstrated that
       no two patients gave the same predicted amino acid sequence for the V3
       loop region. The sequences revealed both European/North American and
       African motifs at the crown of the V3 loop thus indicating a diversity
       of HIV strains in the SW Thames Region of South London. The results show
       that the confirmatory PCR for HIV-1 can be carried out efficiently on
       the same clotted blood specimens as used for routine HIV serology on
       patients undergoing diagnostic evaluation.
 DE    Africa  Amino Acid Sequence  Comparative Study  DNA, Viral/*BLOOD
       Europe  Gene Products, env/CHEMISTRY/GENETICS  Gene Products,
       gag/CHEMISTRY/GENETICS  *Genes, env  *Genes, gag  Human  HIV
       Antibodies/BLOOD  HIV Infections/BLOOD/*DIAGNOSIS/EPIDEMIOLOGY  HIV
       Seropositivity/BLOOD/DIAGNOSIS  HIV-1/GENETICS/*ISOLATION & PURIF
       Molecular Sequence Data  North America  Plasmids  Polymerase Chain
       Reaction/*METHODS  Proviruses/*ISOLATION & PURIF  Risk Factors
       Sensitivity and Specificity  Sequence Homology, Amino Acid  Support,
       Non-U.S. Gov't  Variation (Genetics)  JOURNAL ARTICLE

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