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Updated: Aug 11, 2026
A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Quantification of hepatitis C virus RNA by RT-PCR in comparison to the branched DNA method
Introduction:
Quantification of hepatitis C virus (HCV) serum viremia levels is an important measure to predict and monitor response to interferon-alpha therapy as well as to predict clinical outcome. We were interested to compare the most widely used HCV quantification methods, quantitative RT-PCR (qRT-PCR) and the branched DNA (bDNA) method, with respect to sensitivity and reliability.
Results:
In the present study, 101 serum samples from patients chronically infected with HCV were simultaneously quantified by an in-house reverse transcriptase (RT)-PCR assay and the branched DNA Quantiplex HCV RNA kit 1.0 and 2.0. The concentration of HCV RNA molecules/ml serum ranged from 1.5 x 10(4) to 1.0 x 10(8) as assessed by our quantitative (q)RT-PCR. Serum concentrations of HCV-genotype 1 were significantly higher when measured with bDNA 1.0 compared to bDNA 2.0 and qRT-PCR, whereas measurements by qRT-PCR and bDNA 2.0 were concordant. Measurements of genotypes 2 and 3 by bDNA 2.0 were significantly higher than by bDNA 1.0. Significantly lower measurements were obtained for genotype 3 by qRT-PCR in comparison to bDNA 2.0.
Conclusion:
Despite subtype-specific differences, there is clinically sufficient agreement in measurements between qRT-PCR and bDNA of HCV RNA concentrations. However, with respect to the therapeutic monitoring of the individual patient, kits and methods should not be exchanged.
