多重PCR检测猪瘟病毒、猪细小病毒和猪伪狂犬病病毒的研究——猪瘟病毒、猪细小病毒和猪伪狂犬病病毒多重PCR引物设计

Studies on the Detection of Classical swine fever,Porcine parvovirus infection and Porcine pseudorabies by Multiplex Polymerase Chain Reaction I Primer ——Design of Multiple PCR Detection of CSFV,PPV、PRV

在GeneBank中查出CSFV、PPV、PRV的所有已知序列。对病毒各基因区进行同源性分析 ,确定CSFV的E2 基因 ,PPV的VP2 基因和PRV的 gⅡ基因为各病毒扩增的靶序列。利用DNAsis对上述保守区进行引物设计 ,对设计出来的 3种病毒引物利用DNAstar进行引物二聚体分析 ,以避免引物之间形成稳定的引物二聚体。选出扩增序列为CSFV 2 88bp、PPV575bp和PRV 70 0bp的 3对引物 ,并对每对引物扩增序列的同源性或互补性进行分析。避免它们之间有较高的同源性或互补性 ,从而确定了

Found out the sequences of all different strains of CSFV, PPV and PRV in the GenBank, then analyzed their homology. According to the property of each virus, the conservative of E (2) in CSFV, the conservative of VP (2) in PPV and the conservative of gⅡin PRV was selected for multiplex PCR application. Design these conservatives’ primers by DNAsis system, and use DNAstar system to analyze them, avoiding the formation of steady dimers among them. The specific fragments to be multiplex PCR amplified were 288 bp for CSFV,(575 bp) for PPV and 700 bp for PRV.These specific fragments were firstly analyzed to avoid high homology and complementarity, and the multiplex PCR primers were successfully designed for the three viruses.