Evaluation of an automated, highly sensitive, real-time PCR-based assay (COBAS Ampliprep™/COBAS TaqMan™) for quantification of HCV RNA

Christoph Sarrazin, Anette Dragan, Barbara C. Gärtner, Michael S. Forman, Stella Traver, Stefan Zeuzem, Alexandra Valsamakis

Research output: Contribution to journalArticle

Abstract

Background: Diagnosis of hepatitis C virus (HCV) infection and its therapy is based on qualitative and quantitative measurement of HCV RNA. Objectives: A new assay that employs automated specimen extraction and real-time RT-PCR (COBAS Ampliprep™/COBAS TaqMan™, "CAP/CTM", Roche Diagnostics, Pleasanton, USA) was designed for linear quantification and highly sensitive detection of HCV RNA. Study design: The performance characteristics of CAP/CTM were compared to standard RT-PCR-based COBAS Amplicor Monitor 2.0 (CAM) assay in a multicenter study. Results: The limit of detection of CAP/CTM was 7.4 IU/ml (95% CI 6.2-10.6) and clinical specificity was 99%. The linear range of HCV RNA quantification by CAP/CTM was between 28 and 1.4 × 107 IU/ml, with a correlation coefficient between expected and observed results of >0.99. A fivefold dilution of serum- or plasma-samples showed a linear correlation of HCV RNA levels in undiluted and diluted samples. Analyses of the mean intra- and inter-assay imprecision within the linear range of quantification showed a coefficient of variation of 3% and 3%, respectively. HCV genotypes 1a/b, 2b, 3a, 4, 5 and 6 were equally quantified by the CAP/CTM and CAM assay with mean deviations ranging from -0.29 log10 to 0.32 log10 IU/ml. HCV RNA quantification by CAP/CTM and CAM was highly concordant (correlation coefficient of 0.96). Conclusions: The CAP/CTM assay is a reliable and robust assay for highly sensitive detection and quantification of HCV RNA within a broad linear range.

Original languageEnglish (US)
Pages (from-to)162-168
Number of pages7
JournalJournal of Clinical Virology
Volume43
Issue number2
DOIs
StatePublished - Oct 1 2008

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Keywords

  • Genotype determination
  • Linearity
  • Precision
  • Sensitivity
  • Specificity

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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