       Document 0592
 DOCN  M9610592
 TI    Performance characteristics for the quantitation of plasma HIV-1 RNA
       using branched DNA signal amplification technology.
 DT    9601
 AU    Todd J; Pachl C; White R; Yeghiazarian T; Johnson P; Taylor B; Holodniy
       M; Kern D; Hamren S; Chernoff D; et al; Chiron Corporation, Emeryville,
       CA 94608, USA.
 SO    J Acquir Immune Defic Syndr Hum Retrovirol. 1995;10 Suppl 2:S35-44.
       Unique Identifier : AIDSLINE MED/96033810
 AB    Highly sensitive assays that quantitate human immunodeficiency virus
       type 1 (HIV-1) RNA may be valuable for clinical research and the
       treatment of HIV-1-infected patients. In this study we evaluated the
       reproducibility and accuracy of the first-generation branched DNA
       (bDNA-1.0) signal amplification assay under conditions that are relevant
       to routine use in a clinical context. We show that the bDNA-1.0 assay
       was able to discern two- to three-fold changes in plasma HIV-1 RNA
       levels as significant. Reverse transcription coupled to polymerase chain
       reaction (RT-PCR) was less reproducible and required a 3.7- to 5.8-fold
       change in plasma HIV-1 RNA levels to be statistically significant. The
       accuracy of the bDNA-1.0 assay in RNA quantitation was not affected by
       HIV-1 genotypic variation or by the presence of hemoglobin, bilirubin,
       lipemia, or any of a dozen therapeutic drugs. Using the bDNA-1.0 assay,
       we show that HIV-1 RNA levels in plasma specimens were stable when
       stored at -80 degrees C and were able to withstand at least three
       freeze-thaw cycles without significant loss. We also examined the
       performance of an ultrasensitive bDNA assay with improvements to the
       signal amplification technology. The ultrasensitive bDNA assay displayed
       a quantitation limit of approximately 500 RNA Eq/ml, yet maintained a
       dynamic quantitation range up to 1.6 x 10(6) RNA Eq/ml. Like the
       bDNA-1.0 assay, the ultrasensitive bDNA assay was not affected by HIV-1
       genotype variability.
 DE    Anticoagulants/STANDARDS  Antiviral Agents/PHARMACOLOGY  Bilirubin/BLOOD
       Blood Preservation  DNA, Viral/ANALYSIS  Gene Amplification  Genotype
       Hemolysis  Human  HIV-1/*GENETICS  Lipids/BLOOD  Polymerase Chain
       Reaction  Reproducibility of Results  RNA, Viral/*BLOOD  Sensitivity and
       Specificity  Specimen Handling  Transcription, Genetic  Variation
       (Genetics)  JOURNAL ARTICLE

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

