A Multi-real-time Fluorescence PCR Method Study On HR-HPV Genotyping Test

Objective: To establish a multi-real-time fluorescence PCR method for simultaneous detection of 13 high risk human papillomavirus(HR-HPV),and to provide a new detection kit for clinical screening of cervical cancer.Methods: Primer Express 3.0 software was used to design specific primers and probes for the non-L1 region(including non-L1 regions,such as E6,E7,E1 region and the the whole genome of HR-HPV as the target sequence).Then a multi-real-time fluorescence PCR was established by evaluating its accuracy,sensitivity and specifity.And the method was optimized for further analysis.Results: The method was tested by using the HPVgenotyping reference.And the result showed that the kit had good accuracy,sensitivity and specifity.The detection sensitivity was 500 copies/m L.No HPV-specific amplified signal appeared when tested with other strains.In clinical samples testing,the positive coincidence rate was 96.43%,the negative coincidence rate was 100%,and the total coincidence was 97.22%.Conclusion: The multi-real-time fluoresence PCR method was proved to be an effective method with good accuracy,sensitivity and specifity and could be widely used in clinical screening of cervical cancer.