Establishment and application of multiplex nano-PCR detection method for ASFV,PRV wild virus and PRV vaccine virus
In order to establish a method for simultaneous detection of ASFV,PRV wild virus and PRV vaccine virus(gE-),a multiplex nano-PCR method for simultaneous detection of ASFV,PRV wild virus and PRV vaccine virus were established by using the B646L gene of ASFV and the gE and gD gene of PRV as the detection objects.The method's specificity and sensitivity were studied and the results showed that it could specifically amplify the target genes of ASFV,PRV wild virus and PRV vaccine strain,but the re-suits were negative when detecting Seneca virus,porcine haemagglutinating encephalomyelitis virus,porcine vesicular stomatitis virus and porcine circovirus type 2.The minimum detection limits of B646L gene,gD gene and gE gene were 4,7 and 8 copies/μL,respectively.In addition,the effectiveness of the multiplex nano-PCR detection method was compared with commonly used commercial kits for these viruses,and the results showed that the coincidence rate was 100％.In summary,the multiplex nano-PCR method established in this experiment has high specificity and sensitivity and could simultaneously detect ASFV,wild-type and vaccine PRV,which could be used for differential diagnosis and disease prevention and control of major diseases in pigs.
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