Abstract A single-round real-time polymerase chain reaction
(PCR) assay based on SYBR green dye technology for
the detection and quantification of hepatitis B virus (HBV)
DNA in serum was evaluated and compared with a
qualitative nested PCR and the Cobas Amplicor HBV
Monitor assay (Roche Molecular Diagnostics, Milan, Italy).
The performance of the real-time PCR assay was evaluated
in a routine clinical laboratory setting with a total of 212
clinical specimens. The sensitivity of the real-time PCR
corresponded to 31 IU/ml (70 copies/ml), and comparison
with the qualitative nested PCR showed significant concordance
for 94% of samples. The linear curve over 7 log
units, spanning 103–109 IU/ml (2.28×103 to 2.28×109
copies/ml), was observed in the quantitative determination.
The interexperimental variability coefficient of the assay
ranged from 0.22 to 0.39 and the intraexperimental
variability coefficient from 0.24 to 0.41. By excluding
values outside of the dynamic ranges of both tests, the HBV
Monitor and the real-time PCR gave an agreement within
±1 log unit for 90% of samples, while those for the
remaining 10% were found to be above 1 log unit but less
than 1.5 log units. When the results inside and outside the
dynamic range of the HBV Monitor were examined, 90%
of the results were in agreement. In conclusion, the realtime
PCR based on SYBR green technology proved
suitable for routine diagnostic purposes, showing good
sensitivity, high specificity, high reproducibility, and good
linearity over a broad dynamic range of quantification.