title: Comparative evaluation of three assays for the quantitation of human immunodeficiency virus type 1 RNA in plasma.
abstract: Reverse transcriptase-coupled polymerase chain reaction (Amplicor HIV-1 Monitor), the branched DNA (bDNA) method (Quantiplex HIV-1 RNA) and the nucleic acid sequence-based assay (NASBA HIV-1 RNA QT) were comparatively evaluated for the quantitation of human immunodeficiency virus type 1 (HIV-1) RNA in plasma. Among 60 plasma specimens from HIV-1 infected patients, HIV-1 RNA was detected in 56 by Amplicor (sensitivity, 93.3%), in 41 by bDNA (sensitivity, 68.3%), and in 60 by NASBA (sensitivity, 100%). HIV-1 RNA was not detected by any of these methods in 34/34 plasma specimens from HIV-1-seronegative blood donors (specificity, 100%). The HIV-1 RNA levels as determined by the different methods were correlated significantly. The frequency of concordant results (log difference < 0.50) was 80.4% between Amplicor and NASBA, 77.5% between Amplicor and bDNA, and 58.6% between bDNA and NASBA. After initiation of antiviral therapy, HIV-1 RNA level variations observed with the three methods were similar. HIV-1 RNA levels were inversely correlated with the CD4+ T cell counts, whereas no correlation was found with HIV-1 p24-antigen levels. When the methods were evaluated for reproducibility, coefficients of variation ranged from 11% to 40% for Amplicor, from 6% to 35% for bDNA, and from 13% to 62% for NASBA. Quantitation of HIV-1 RNA in culture supernatants from HIV-1 subtype A to H strains showed that bDNA can be used to quantitate RNA from all HIV-1 subtypes, whereas Amplicor failed to detect RNA from subtype A strains and NASBA subtype G strains.
authors.affiliation: Etablissement de Transfusion Sanguine Languedoc-Roussillon, Centre Hospitalier Universitaire, Montpellier, France.
authors.forename: J
authors.initials: J
authors.lastname: Coste
authors.forename: B
authors.initials: B
authors.lastname: Montes
authors.forename: J
authors.initials: J
authors.lastname: Reynes
authors.forename: M
authors.initials: M
authors.lastname: Peeters
authors.forename: C
authors.initials: C
authors.lastname: Segarra
authors.forename: J P
authors.initials: JP
authors.lastname: Vendrell
authors.forename: E
authors.initials: E
authors.lastname: Delaporte
authors.forename: M
authors.initials: M
authors.lastname: Segondy
mesh_terms: D018791:CD4 Lymphocyte Count; D005069:Evaluation Studies as Topic; D005821:Genetic Techniques; D016655:HIV Core Protein p24 / Q000097:blood; D015658:HIV Infections / Q000145:classification / Q000821:virology*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014766:Viremia / Q000821:virology; D014773:Virology / Q000379:methods*
chemical_list: D016655:HIV Core Protein p24; D012367:RNA, Viral