TY - JOUR
TI - Development and assessment of a novel real-time PCR assay for
quantitation of HBV DNA
AU - Paraskevis, D
AU - Haida, C
AU - Tassopoulos, N
AU - Raptopoulou, M and
AU - Tsantoulas, D
AU - Papachristou, H
AU - Sypsa, V
AU - Hatzakis, A
JO - Journal of Virological Methods
PY - 2002
VL - 103
TODO - 2
SP - 201-212
PB - ELSEVIER SCIENCE BV
SN - 0166-0934
TODO - 10.1016/S0166-0934(02)00033-2
TODO - HBV; real-time PCR; quantitation
TODO - HBV DNA quantitation is used extensively for the monitoring of treatment
of hepatitis B virus (HBV) infection. The aim of this study was to
develop a highly sensitive and reproducible real-time PCR (RTD-PCR)
assay for the quantitation of HBV DNA using the LightCycler system. The
performance of this assay was assessed by analyzing serial dilutions of
HBV genomic DNA of known concentration and the lower limit of detection
was found to be I DNA copy/reaction. By using serial dilutions of
plasmid standard, RTD-PCR was determined to quantify HBV DNA in a
10-log(10) dynamic range. RTD-PCR was found to be more sensitive than
the commercially available tests such as the Quantiplex(TM) HBV DNA and
the AMPLICOR HBV MONITOR(TM) assays. The median coefficient of variation
of interexperimental variability was 3.2%. The HBV DNA values obtained
with RTD-PCR were highly correlated with assays available commercially.
These findings suggest that our RTD-PCR assay combines high sensitivity
and reproducibility for HBV DNA quantitation in an incomparable high
dynamic range of quantitation. (C) 2002 Published by Elsevier Science
B.V.
ER -