title: An enhanced-sensitivity branched-DNA assay for quantification of human immunodeficiency virus type 1 RNA in plasma.
abstract: The quantification of human immunodeficiency virus type 1 (HIV-1) RNA has facilitated clinical research and expedited the development of antiretroviral drugs. The branched-DNA (bDNA) assay provides a reliable method for the quantification of HIV-1 RNA in human plasma and is considered one of the most reproducible assays ready for use in clinical trials. A series of oligonucleotide probe design and solution changes have been developed to enhance the sensitivity of the bDNA assay while maintaining its performance characteristics. Among the changes incorporated into the enhanced-sensitivity bDNA (ES bDNA) assay to reduce the background level and enhance the signal are the use of shorter overhang sequences of target probes for capture, the cruciform design of target probes for amplification, and the addition of preamplifier molecules. The ES bDNA assay is at least 20-fold more sensitive than the first-generation bDNA assay, yet it maintains a high level of accuracy, linearity, and reproducibility. Further, quantification values obtained with the ES bDNA assay and the first-generation bDNA assay are highly correlated, thus allowing for meaningful comparisons of HIV-1 RNA levels in specimens tested with either assay. The ES bDNA assay may be useful in determining the prognostic value of HIV-1 RNA levels of below 10,000 copies per ml and in assessing the clinical benefit of antiretroviral therapy-induced decreases in plasma HIV-1 RNA sustained at levels of below 10,000 copies per ml.
authors.affiliation: Chiron Corporation, Emeryville, California 94608-2916, USA.
authors.forename: D
authors.initials: D
authors.lastname: Kern
authors.forename: M
authors.initials: M
authors.lastname: Collins
authors.forename: T
authors.initials: T
authors.lastname: Fultz
authors.forename: J
authors.initials: J
authors.lastname: Detmer
authors.forename: S
authors.initials: S
authors.lastname: Hamren
authors.forename: J J
authors.initials: JJ
authors.lastname: Peterkin
authors.forename: P
authors.initials: P
authors.lastname: Sheridan
authors.forename: M
authors.initials: M
authors.lastname: Urdea
authors.forename: R
authors.initials: R
authors.lastname: White
authors.forename: T
authors.initials: T
authors.lastname: Yeghiazarian
authors.forename: J
authors.initials: J
authors.lastname: Todd
mesh_terms: D019380:Anti-HIV Agents / Q000627:therapeutic use; D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D005069:Evaluation Studies as Topic; D014644:Genetic Variation; D005838:Genotype; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology; D017320:HIV Protease Inhibitors / Q000627:therapeutic use; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007546:Isoquinolines / Q000627:therapeutic use; D015336:Molecular Probe Techniques / Q000706:statistics & numerical data; D008969:Molecular Sequence Data; D019888:Nelfinavir; D021141:Nucleic Acid Amplification Techniques; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013451:Sulfonic Acids / Q000627:therapeutic use; D014773:Virology / Q000379:methods* / Q000706:statistics & numerical data
chemical_list: D019380:Anti-HIV Agents; D015342:DNA Probes; D017320:HIV Protease Inhibitors; D007546:Isoquinolines; D012367:RNA, Viral; D013451:Sulfonic Acids; D019888:Nelfinavir