Quantification of hepatitis C virus RNA by RT-PCR in comparison to the branched DNA method.
BACKGROUND
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.


CONCLUSIONS
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.
Roth
W K
WK
Georg-Speyer-Haus, Frankfurt am Main, Germany.
Lee
J H
JH
Rüster
B
B
Zeuzem
S
S
D004279:DNA, Viral / Q000097:blood* / Q000235:genetics; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique; D012680:Sensitivity and Specificity; D014766:Viremia / Q000175:diagnosis* / Q000821:virology
D004279:DNA, Viral; D012367:RNA, Viral