多重PCR快速诊断猪伪狂犬病毒、猪圆环病毒2型与猪细小病毒方法的建立

Establishment of multiplex PCR for rapid detection of porcine pseudorabies virus,porcine circovirus type Ⅱ and porcine parvovirus

利用Primer Premier 5.0及DNAstar软件对猪伪狂犬病毒gB基因、猪圆环病毒2型ORF2基因和猪细小病毒VP2基因的保守区进行引物设计,选出扩增序列为猪伪狂犬病毒(373 bp)、猪圆环病毒2型(430 bp)和猪细小病毒(495 bp)的3对引物.敏感性和特异性试验结果表明,mPCR对3种病毒的核酸检测限量从猪伪狂犬病毒至猪细小病毒分别为1.62×10-6、1.47×10-6和1.28×10-4ng,对灭菌双蒸水、猪繁殖与呼吸障碍综合征病毒cDNA和猪瘟病毒cDNA的mPCR扩增结果均为阴性.mPCR可作为临床上猪伪狂犬病、猪圆环病毒2型感染和猪细小病毒病的病原学快速诊断方法.

According to the gene sequences in C, enBank of PRV gB, PCV20RF2, PPV VP2, specific primers for each of three common DNA viruses were designed by Primer Premier 5.0 and DNAstar software, and three specific bands, respectively, PRV （373 bp）, PCV2（430 bp）, PPV（495 bp） were amplified. It was shown that mPCR was able to detect as little as 1.62 ×10-6, 1.47 × 10-6 and 1.28 × 10-4 ng of each viral target through the highly sensitive and specificity assay. And detections of ddH2O, PRRSV, SV were negative at the same time. In conclusion, mPCR would be useful in routine molecular diagnosis and epidemiology of PRV, PCV2 and PPV.