       Document 0951
 DOCN  M9650951
 TI    Quantitation of human immunodeficiency virus type 1 DNA by two PCR
       procedures coupled with enzyme-linked oligosorbent assay.
 DT    9505
 AU    Mallet F; Hebrard C; Livrozet JM; Lees O; Tron F; Touraine JL; Mandrand
       B; Unite Mixte de Recherche 103 Centre National de la Recherche;
       Scientifique-bioMerieux, Ecole Normale Superieure de Lyon,; France.
 SO    J Clin Microbiol. 1995 Dec;33(12):3201-8. Unique Identifier : AIDSLINE
       MED/96156129
 AB    Two quantitative PCR methods with our nonisotopic enzyme-linked
       oligosorbent assay (ELOSA) in microtiter plate format were developed for
       quantitation of human immunodeficiency virus type 1 (HIV-1).
       Quantitative competitive PCR (QC-PCR) was based on the coamplification
       of the wild-type nef region with a mimic competitive nef gene template
       carrying mutations in the capture region. Correlation of wild-type HIV-1
       nef DNA to mimic template copy number permitted quantitation of HIV-1
       copy numbers in the range of 20 to 2,000 copies per micrograms of DNA.
       Internally controlled PCR (IC-PCR) was based on coamplification of the
       nef region and the ras gene as an internal endogenous standard.
       Correlation to known amounts of HIV-1 DNA permitted quantitation by
       IC-PCR of HIV-1 copy numbers in the range of 10 to 2,000 copies per
       microgram of DNA. QC- and IC-PCR-ELOSA were performed on a panel of 53
       seropositive patients and 12 seronegative controls. The methods showed
       similar coefficients of variation below 24%. Quantitations by QC- and
       IC-PCR-ELOSA were identical for 77% of patient samples. The copy level
       ranged between 443 +/- 156 and 21,453 +/- 13,511 copies per 10(5) CD4
       cells for asymptomatic and AIDS patients, respectively. The simplicity
       and reliability of QC- and IC-PCR-ELOSA methods make them appropriate
       for routine laboratory use in the quantitation of viral and bacterial
       DNAs.
 DE    Acquired Immunodeficiency Syndrome/VIROLOGY  Base Sequence  Comparative
       Study  DNA Primers/GENETICS  DNA, Viral/*ANALYSIS/*GENETICS
       Enzyme-Linked Immunosorbent Assay/*METHODS/STATISTICS & NUMER
       DATA/STANDARDS  Evaluation Studies  Genes, nef  Human  HIV
       Infections/VIROLOGY  HIV-1/*GENETICS/*ISOLATION & PURIF  Indicators and
       Reagents  Molecular Sequence Data  Polymerase Chain
       Reaction/*METHODS/STATISTICS & NUMER DATA  Quality Control
       Reproducibility of Results  JOURNAL ARTICLE

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

