Abstract The quantification of viral
nucleic acids in
serum by
real-time PCR plays an important
role in diagnosing
hepatitis B virus and
hepatitis C virus infection. In this study, we developed an assay using specific primers and probes to quantify
hepatitis B virus DNA or
hepatitis C virus RNA in
serum from infected
patients. For
standardization and validation of the assay, an international panel of
hepatitis B virus/
hepatitis C virus and standard
plasmids was used. A correlation coefficient of 0.983 and 0.963 for
hepatitis B virus and
hepatitis C virus, respectively, was obtained based on cycle threshold values and concentrations of
DNA or
RNA. The standard curve showed a linear relationship from 19 IU/mL to 1.9 × 109 IU/mL of
serum, with a coefficient of
determination (r2) of 0.99. In sera from
patients infected with
hepatitis B virus or
hepatitis C virus viral loads (19 IU/mL and 1.9 × 109 IU/mL), we quantified viral loads with a
detection limit of 1.9 × 102 IU/mL. The real-
time quantitative
PCR assay developed in this study provides an ideal system for routine
diagnosis and confirmation of indeterminate serological results, especially in immunosuppressed
patients.