RT Journal Article
SR Electronic
T1 Detection of PCR products using self-reporting duplex mutation primers
JF Journal of Clinical Pathology
JO J Clin Pathol
FD BMJ Publishing Group Ltd and Association of Clinical Pathologists
SP 1046
OP 1048
DO 10.1136/jcp.2009.064402
VO 62
IS 11
A1 Q-F Xia
A1 Z-G Tu
A1 J-B Liu
A1 X Qin
A1 S-Y Qian
A1 P Li
YR 2009
UL http://jcp.bmj.com/content/62/11/1046.abstract
AB 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.