To establish a rapid detection and identification method for foot-and-mouth disease virus type-O(FMDV-O)and Seneca Valley virus(SVV),primers and probes were designed based on the VP2 gene of FMDV-O and the P3 gene of SVV.After optimizing the reaction conditions,a duplex real-time RT-PCR method was established for simultaneous detection and identification of FMDV-O and SVV.The results showed that this method has strong specificity and can detect the VP2 gene of FMDV-O and P3 gene of SVV simultaneously without cross-reaction to other viral nucleic acids,including vesicular stomatitis virus(VSV),swine vesicular disease virus(SVDV),classical swine fever virus(CSFV),porcine reproductive and respiratory syndrome virus(PRRSV),porcine parvovirus(PPV),foot-and-mouth disease virus type-A(FMDV-A)and foot-and-mouth disease virus type-A-1(FMDV-A-1)nucleic acids.The method exhibited broad-spectrum coverage of epidemic strains of foot-and-mouth disease virus type-O.The method demonstrated good sensitivity,and the detection limit for the plasmid standard could reach about 7.17 × 102 copies/μL in this study.It also exhibited good repeatability with a variation coefficient of Ct values of less than 3％.The method established in this experiment was used to detect 30 clinical samples,and the results were consistent with those of the reference method.In conclusion,this method enables rapid and accurate identification of FMDV-O and S VV,creating technical conditions for efficient quarantine at ports of entry.
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