Detection of PCR products using self-reporting duplex mutation primers
- 1Faculty of Laboratory Medicine, Hainan Medical College, Haikou, PR China
- 2Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing, PR China
- Correspondence to Dr Q-F Xia, Faculty of Laboratory Medicine, Hainan Medical College, Haikou, 570102, PR China; xiaqianfeng{at}sina.com
- Accepted 22 June 2009
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.
Footnotes
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Funding This work was supported by grant HjKj200524 from the Education Bureau of Hainan province.
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Competing interests None.
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Ethics approval Obtained.
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Provenance and peer review Not commissioned; not externally peer reviewed.
