Development And Clinical Application Of A Novel Fluorescent Quantitative PCR Detection Method Of Hepatitis B Virus

Objective:As HBV has been detected by fluorescent quantitative PCR methods, which need long time and high price, we tended to establish a new quickly real-time fluorescence PCR method to detect hepatitis B virus (HBV) with high sensitivity, specificity, stability and lower price using the duplex scorpion primer.Method:The duplex scorpion primers were designed according to a settled nucleic acid sequence of hepatitis B virus. The targeted sequence was cloned into the vector pMD18-T. The recombinant vector was used as standard template. After optimization the PCR reaction system and method evaluation, the new real time fluorescence PCR method was used to detect clinic examples comparing with the commercial Taqman-FQ-PCR.Results: 1) Restriction maps showed that the structure of recombinant plasmid was the same as expected. 2) Methodology analysis showed that the duplex scorpion primer method was successfully established with high sensitivity, specificity and accuracy. 3) After applied in clinical experiments, this new fluorescent quantitative method has many...