Multiplex PCR for rapid detection of porcine circovirus type II,porcine parvovirus,porcine pseudorabies virus and porcine reproductive and respiratory syndrome virus

A multiplex PCR(mPCR) assay was developed and subsequently evaluated for its effectiveness as a means to simultaneously detect multiple viral infections of swine.Specific primers for each of four common DNA and RNA viruses were designed by Oligo 6.0 software,namely,porcine circovirus type Ⅱ(PCV2 ORF2),porcine parvovirus(PPV NS-1),pseudorabies virus(PRV gB),porcine reproductive and respiratory syndrome virus(PRRSV MN),four specific bands,respectively,353 bp(PCV2),271 bp(PPV),434 bp(PRRSV) and 194 bp(PRV),were amplified.The assay was shown to be highly sensitive and specificity when detect composite of all four virus by purpose.It was also effective for detecting one or more of these four same viruses in various combinations in specimens including lymph nodes,lungs,spleens,and tonsils collected from clinically ill pigs,and aborted fetuses.The relative efficiency(compared to performing separate assays for each virus) and apparent sensitivity of mPCR suggest its potential application for routine molecular diagnostic purposes and investigation epidemiology.