Establishment and application of a mutiplex PCR diagnosis method for reproductive failure of swine diseases

According to the sequences published in the GenBank,the sequences of relevant genes of swine fever virus (CSFV),porcine circovirus type 2 (PCV2),porcine reproductive and respiratory syndrome virus (PRRSV) and porcine pseudorabies virus (PRV) were analyzed by biological softwares such as DNAStar and BLAST,and four pairs of special primers were designed by the Oligo 6.0 software.On the basis of initial optimization of individual PCR reaction annealing temperature and the reaction system,a multiple PCR diagnostic method for simultaneously detecting the four viruses of CSFV,PCV2,PRRSV and PRV was established.And then,this method was used for detection of 30 clinical samples.The results showed that positive rate of CSFV,PRRSV,PCV2,PRV were 46.7 %,43.3 %,3.4 %,10 % respectively.Among them,the positive rate of PRV-PCV2,CSFV-PCV2 and CSFV-PRRSV co-infection were 6.7 %,36.7 % and 10 % respectively,while positive rate of PRV,PCV2 and PRRSV single infection was 3.3 %,10 % and 33.3 % respectively.The results showed that the multiplex PCR had good specificity,sensitivity,reproducibility,and could be used in clinical differential diagnosis.