Abstract

A novel oligonucleotide probe, with duplex mutation primers, which comprised a normal reverse primer labelled with a fluorophore at its 5′-end and a single-base mismatched complementary oligonucleotide labelled with a quencher at its 3′-end, was designed. Its application in the detection of hepatitis B virus (HBV) DNA of serum was investigated; results were compared with those obtained using a commercial TaqMan kit. There was a good linear correlation in the range of 10²∼10⁷ copies/ml (r² = 0.999) with the method. Intra-experimental coefficients of variation (CVs) were 0.70%∼7.80%, and inter-experimental CVs were 0.78%∼9.02%, respectively. There was no significant difference of HBV genome number tested by the two methods (p<0.05) in 132 hepatitis B patients; HBV DNA was not detected in any serum samples of 20 healthy volunteers by the two methods.