Multiplex PCR for rapid detection of pseudorabies virus,porcine parvovirus and porcine circovirus type 2

A multiplex PCR(mPCR) assay was developed and evaluated for its effectiveness as a means to simultaneously detect multiple viral infection of swine . Specific primers for each of the three common DNA viruses, Pseudorabies virus(PRV) , Porcine parvovirus ( PPV) and Porcine circovirus type 2(PCV2) were used to test the procedure, Four specific bands of 269bp(PCV2), 583 bp(PPV) 372 bp(PRV gB)and147 bp(PRV gE)were amplified. The assay proved to be sensitive when a composite of all three viruses were amplified, including both field and gene-deleted permutations of PRV. No specific band was amplified from other pathogenic viruses and bacteria. As little as 10 pg PCV2, 10-6.2 TCID50 PPV,10-3.8 TCID50\PRV gB and 10-5.8 TCID50 PRV gE were detected in this mPCR. This method could effectively detect infection of PCV2, PPV, field and gene-deleted permutations of PRV from clinical samples.