Detection of porcine pseudorabies virus and differentiation of natural PRV from vaccinal PRV by multi-PCR

A multiplex polymerase chain reaction was developed and optimized to detect simultaneously porcine pseudorabies virus (PRV) and differentiate natural PRV from vaccinal TK~(-)/gI~(-)/gE~(-)PRV. Three pairs of specific primers were designed according tothe conservative sequences of PRV gB,gD and gE genes in GenBank. All samples containing PRV and recombinant PRV were amplified desirably by the multiplex PCR using the three sets of primers and then the multiplex PCR conditions were optimized. Three expected specific bands of PRV gB gene 549bp, gD gene 429bp and gE gene 366bp in length with the templates as little as 756pg of PRV or 106pg of vaccinal TK~(-)/gI~(-)/gE~(-) PRV were obtained by the multiplex PCR.The specific experiment showed that the multiplex PCR is qualified specifically to detect natural PRV and vaccinal TK~(-)/gI~(-)/gE~(-)recombinant PRV.