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Detection of PCR products using self-reporting duplex mutation primers
  1. Q-F Xia1,
  2. Z-G Tu2,
  3. J-B Liu2,
  4. X Qin1,
  5. S-Y Qian1,
  6. P Li1
  1. 1
    Faculty of Laboratory Medicine, Hainan Medical College, Haikou, PR China
  2. 2
    Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing, PR China
  1. Correspondence to Dr Q-F Xia, Faculty of Laboratory Medicine, Hainan Medical College, Haikou, 570102, PR China; xiaqianfeng{at}sina.com

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.

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Footnotes

  • Funding This work was supported by grant HjKj200524 from the Education Bureau of Hainan province.

  • Competing interests None.

  • Ethics approval Obtained.

  • Provenance and peer review Not commissioned; not externally peer reviewed.

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