Establishment and Application of a Multiplex PCR Method for Single-step Simultaneous Detection of Six Viral Pathogens of Porcine Reproductive Disorder

According to the gene sequences in GenBank,six pairs of specific primers were designed to amplify part fragments of the specific genes of PRV gE,PPV NS1,PCV ORF2,PRRSV ORF7,CSFV E2 and JEV E,respectively.The results of sensitivity and specificity tests showed that the minimum amounts of nucleic acid detection by the mPCR were as follows: PRV 6.6 pg,PPV 96 pg,PCV2 12.9 pg,PRRSV 10.5 pg,CSFV 51 pg,and JEV 46 pg respectively,and the amplification results of Swine Influenza Virus(SIV) and Escherichia coli(E.coli) were both negative.The results of 103 clinical samples from Guizhou province revealed the six viral diseases exist universally in pig farms of Guizhou province,and the mixed infection was much serious.The Six-plex PCR method could detect the infection of PRV,PPV,PCV,PRRSV,CSFV and JEV simultaneously,could detect DNA and RNA virus synchronously,and could differentiate and diagnose the clinical samples of single infection or mixed infection infected by the six viruses quickly.