Detection of hepatitis B virus by fluorescence quantitative PCR
Objective To set up a Fluorescence quatitative PCR assay(FQ PCR) to detect HBV DNA and to compare the detected results with those from agarose gel electrophoresis/EB staining. Methods A pair of primers and one probe with 2 fluorescent groups were synthesized. The FQ PCR was carried out in Gene Amp5700 Sequence Detection System followed by agarose gel electrophoresis/EB staining detection of the PCR products. Results A quantitative fluorescence PCR assay for detection of HBV DNA was set up. A standard curve was made ploting 3 to 4 concentrations of known positive HBV DNA as template. 698 serum samples were detected by the FQ PCR with positive rate 29.2%(204/698). A 314bp band was identified in 193 samples out of the 698 cases (27.7%). Conclusion The FQ PCR assay for HBV showed high sensitivity, good specificity, more automation without cross contamination and so can be used to detect HBV DNA accurately and quantitatively. [