title: Rapid and precise quantification of HIV-1 RNA in plasma using a branched DNA signal amplification assay.
abstract: The level of human immunodeficiency virus type 1 (HIV-1) RNA in human plasma has been quantitated directly with use of a solid-phase nucleic acid hybridization assay, based on branched DNA (bDNA) signal amplification technology with chemiluminescent detection. Signal amplification is accomplished by the incorporation of sites for 1,755 alkaline phosphatase-labeled probes per genome of HIV-1, after successive hybridization of target-specific oligonucleotides and bDNA amplifier molecules. The assay is performed in microwells, much like an immunoassay, and is amenable to routine laboratory use. Reproducibility and specificity studies indicated that the bDNA method was precise and showed no reactivity with seronegative donors. HIV-1 RNA levels were quantitated for 348 seropositive specimens, with a detection rate of 83% for those specimens from patients with < 500 CD4+ T-cell counts. Plasma RNA levels were found to change with disease stage, and in response to antiviral therapy. Quantitation of HIV-1 RNA in the plasma of HIV-1-infected patients, with use of the bDNA assay, may be a useful method for monitoring HIV-1 disease progression and therapeutic response.
authors.affiliation: Nucleic Acid Systems, Chiron Corporation, Emeryville, California 94608, USA.
authors.forename: C
authors.initials: C
authors.lastname: Pachl
authors.forename: J A
authors.initials: JA
authors.lastname: Todd
authors.forename: D G
authors.initials: DG
authors.lastname: Kern
authors.forename: P J
authors.initials: PJ
authors.lastname: Sheridan
authors.forename: S J
authors.initials: SJ
authors.lastname: Fong
authors.forename: M
authors.initials: M
authors.lastname: Stempien
authors.forename: B
authors.initials: B
authors.lastname: Hoo
authors.forename: D
authors.initials: D
authors.lastname: Besemer
authors.forename: T
authors.initials: T
authors.lastname: Yeghiazarian
authors.forename: B
authors.initials: B
authors.lastname: Irvine
mesh_terms: D000212:Acyclovir / Q000627:therapeutic use; D018791:CD4 Lymphocyte Count; D004277:DNA, Single-Stranded; D016049:Didanosine / Q000627:therapeutic use; D018450:Disease Progression; D004359:Drug Therapy, Combination; D005784:Gene Amplification; D015751:Genes, pol; D006679:HIV Seropositivity / Q000175:diagnosis / Q000188:drug therapy / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008137:Longitudinal Studies; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012367:RNA, Viral / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015215:Zidovudine / Q000627:therapeutic use
chemical_list: D004277:DNA, Single-Stranded; D015345:Oligonucleotide Probes; D012367:RNA, Viral; D015215:Zidovudine; D016049:Didanosine; D000212:Acyclovir