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title£ºEstablishment and application of a multiplex PCR method for detection of six swine virus DNA (PRV,PPV,PCV2) and RNA (PRRSV,CSFV,JEV) simultaneously Abstract: According to the gene sequences in GenBank, six pairs of specific primers were designed to amplify fragments of the specific genes of PRV gE, PPV NS1, PCV2 ORF2, PRRSV ORF7, CSFV E2 and JEV E, respectively. The results of the multiplex PCR showed that the minimum nucleic acid detection amount of PRV, PPV, PCV2, PRRSV, CSFV and JEV were 6.6pg, 96pg, 12.9pg, 10.5pg, 51pg and 46pg, respectively, and both Swine Influenza Virus (SIV) and Escherichia coli (E.coli) were negative. The results of 103 clinical samples from Guizhou province revealed that the six viruses were positive in all pig farms in Guizhou province, and mixed infections were possible. The six-plex PCR method not only could simultaneously detect the infection of PRV, PPV, PCV, PRRSV, CSFV and JEV, but also could detect DNA and RNA virus synchronously, and could rapidly differentiate and diagnose the clinical samples of single infection or mixed infections of the six viruses. ÏȺóͶÁËÁ½¸öÔÓÖ¾£¬¶¼Òò²»¶Ô¿Ú£¬ÌØÇóÖúÏ£ÍûÄÜÍÆ¼ö¼¸¸öºÏÊʵÄSCIÆÚ¿¯¡£Ð»Ð»£¡ |
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