Development And Application Of A Duplex Real-Time RT-PCR Assay For Detection Of GX And XJ Strain Of Porcine Type O FMDV

To developing a duplex real-time RT-PCR method for detecting different genotypes from type O foot-and-mouth disease viruses (FMDV), one pair of universal primers and two Taqman probes were designed and synthesized according to the VP1 gene sequences of FMDV O/GX/09-7 and O/XJ/10-11 strain. Through preparing of GX and XJ strain standard RNA, then optimizing reaction conditions, the duplex real-time RT-PCR will be developed in this study. And its sensitivity, specificity and repeatability will be evaluated. The inactivated porcine type O FMDV GX and XJ strain will be detected Using the developed duplex real-time RT-PCR in this study.The results showed that the correlation coefficients of duplex real-time standard curve for GX and XJ strain were R2=0.997 and R2=0.995, respectively. This developed duplex real-time RT-PCR method with good specificity and sensitivity can identify different genotypes of FMDV in inactivated virus, and the lowest detection limit of RNA was 10 copies/μL. This study can be used for virus in purified inspection and also provided a new detection method for quality control of FMDV vaccine production.