【摘要】 为建立猪群常见疫病多重PCR诊断方法,本研究据GenBank发表的猪瘟病毒(classical swine fever virus,CSFV)、猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)、猪伪狂犬病病毒(pseudorabies virus,PRV)、猪圆环病毒2型(porcine circovirus type 2,PCV2)和猪细小病毒(porcine parvovirus,PPV)基因序列,设计并合成了5对特异性引物。通过反应条件优化、特异性、敏感性和重复性测定,建立了能同时扩增CSFV、PRRSV、PRV、PCV2、PPV5种病毒的多重PCR方法。结果表明该方法只能检测出CSFV、PRRSV、PRV、PCV2、PPV 5种病毒,不能检测出猪流感病毒(swine influenza virus,SIV)、猪乙型脑炎病毒(Japanese encephalitis virus,JEV)、猪链球菌(Streptococcus suis,SS)和猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV),能检测到CSFV、PRRSV、PRV、PCV2和PPV的核酸浓度分别为220、1.6、72、400和370pg。初步应用结果表明,建立的多重PCR方法适用于对CSFV、PRRSV、PRV、PCV2和PPV的快速诊断。更多还原

【Abstract】 To establish a method for simultaneous detection of classical swine fever virus(CSFV),porcine reproductive and respiratory syndrome virus(PRRSV),pseudorabies virus(PRV),porcine circovirus type 2(PCV2) and porcine parvovirus(PPV),a multiplex PCR was developed with a set of specific primers designed based on the conserved sequences of CSFV,PRRSV,PRV,PCV2 and PPV.Under the optimized conditions of multiplex PCR,five special fragments of 167(CSFV),433(PRRSV),305(PRV),559(PCV2)and 882bp(PPV)were amplified with a detection limit of 220,1.6,72,400 and 370pg,respectively.But the multiplex PCR amplification results of swine influenza virus(SIV),Japanese encephalitis virus(JEV),Streptococcus suis(SS)and porcine epidemic diarrhea virus(PEDV)were negative.The results showed that the multiplex PCR method was capable of CSFV,PRV,PRRSV,PCV2,PPV infection of single or mixed clinical samples for rapid diagnosis.更多还原