Detection of PCR products using self-reporting duplex mutation primers

作者：Q-F Xia, Z-G Tu, J-B Liu, Xi Qin, S-Y Qian, Pingping Li · 发表于：Journal of Clinical Pathology · 年份：2009 · DOI：10.1136/jcp.2009.064402 · 被引用次数：2 · 研究领域：Hepatitis B Virus Studies、Viral gastroenteritis research and epidemiology、Bacteriophages and microbial interactions

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 10(2) approximately 10(7) copies/ml (r(2) = 0.999) with the method. Intra-experimental coefficients of variation (CVs) were 0.70% approximately 7.80%, and inter-experimental CVs were 0.78% approximately 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.