Multiplex PCR Assay for Detection of Porcine Parvovirus,Pseudorabies Virus and Porcine Circovirus Type

Based on the similarity of the viral sequences deposited in the GenBank database,the VP2 gene of PPV,the gB gene of PRV,and the ORF1 gene of PCV2 were selected as the diagnostic targets.By using three pairs of virus-specific primers,three PCR assay were established to amplify the conservative regions of the three viruses,respectively.Consequently,a multiplex PCR method to detect the three viruses in one tube was developed.The multiple PCR system would amplify a 203 bp segments for PPV,a 368 bp segments for PRV and a 703 bp segments for PCV2 simultaneously or separately in the samples,depending on its infection status.To evaluate the multiplex PCR assay,78 clinical samples were comparatively detected.The data showed that the multiple PCR method being 97.4 coincidence with the single PCR.The results indicated that the optimized multiplex PCR is both sensitive and specific,providing a new method for the clinical diagnosis and epidemiological research of these three diseases.