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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.

Original publication

DOI

10.1128/jcm.34.12.3196-3202.1996

Type

Journal article

Journal

Journal of clinical microbiology

Publication Date

12/1996

Volume

34

Pages

3196 - 3202

Addresses

Chiron Corporation, Emeryville, California 94608-2916, USA.

Keywords

Humans, HIV-1, HIV Infections, Sulfonic Acids, Isoquinolines, Nelfinavir, DNA Probes, RNA, Viral, HIV Protease Inhibitors, Anti-HIV Agents, Sensitivity and Specificity, Reproducibility of Results, Molecular Probe Techniques, Nucleic Acid Amplification Techniques, Virology, Base Sequence, Genotype, Molecular Sequence Data, Evaluation Studies as Topic, Genetic Variation