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Performance characteristics of a real-time RT-PCR assay for quantification of hepatitis C virus RNA in patients with genotype 1 and 2 infections
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Jee-Fu Huang
Published/Copyright:
April 1, 2008
Abstract
Background: Polymerase chain reaction (PCR) methods play an essential role in providing data relating to diagnosis, monitoring and treatment of hepatitis C virus (HCV) infection. The real-time reverse transcription PCR (RT-PCR) assay is an established and promising tool in terms of quantifying HCV RNA for clinical application. This study aimed to evaluate the performance characteristics of a real-time RT-PCR-based test in a clinical setting.
Methods: Validation and reproducibility tests were performed using a standard panel. Sera from 197 chronic HCV patients were analyzed by the real-time RT-PCR assay and the results were compared with the Versant bDNA3.0 assay (bDNA3.0).
Results: The real-time RT-PCR assay showed an acceptable linear response (r2=0.989–0.995) in the serial dilutions regarding genotypes 1b, 2a, 2b and 1b+2a. HCV viral loads were quantifiable in all 197 patients (100%) by the real-time RT-PCR assay and in 194 (98.5%) by the bDNA3.0. HCV RNA quantification values measured by the real-time RT-PCR and bDNA3.0 assays were positively correlated (Pearson's correlation coefficient r=0.734, p<0.001). The real-time RT-PCR assay values were on average 0.13 logs higher than the bDNA3.0 results. The correlation coefficients with genotypes 1b, 2a, 2b and mixed were 0.737, 0.711, 0.791 and 0.766, respectively (p<0.01).
Conclusions: The real-time RT-PCR assay showed comparable performance with bDNA3.0 regarding quantification of HCV viral loads with genotype 1 and 2 HCV infections.
Clin Chem Lab Med 2008;46:475–80.
Keywords: hepatitis C virus; quantification assays; real-time reverse transcription polymerase chain reaction assay (RT-PCR)
Received: 2007-10-2
Accepted: 2007-11-27
Published Online: 2008-04-01
Published in Print: 2008-04-01
©2008 by Walter de Gruyter Berlin New York
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Keywords for this article
hepatitis C virus;
quantification assays;
real-time reverse transcription polymerase chain reaction assay (RT-PCR)
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- Certification of reference materials for detection of the human prothrombin gene G20210A sequence variant
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- A new single nucleotide polymorphism genotyping method based on gap ligase chain reaction and a microsphere detection assay
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