多重RT-PCR快速检测鉴别新城疫病毒强毒株和弱毒疫苗株方法的建立

根据新城疫病毒(NDV)基因结构的特点及强、弱毒株融合基因裂解位点的序列差异设计了4条引物,建立了一种可以快速鉴别NDV强毒株和弱毒疫苗株的多重PCR方法。检测结果显示,强毒株可以扩增出442 bp的特异性片段和671 bp的通用片段,弱毒疫苗株可以扩增出252bp的特异性片段和671 bp的通用片段。该方法只需进行一次RT-PCR,整个过程可在数小时内完成。经敏感性测定,该方法最低能检测到100 pg的NDV RNA。

Development of multiplex RT-PCR method for rapid differentiation of NDV virulent strains from avirulent strains

Four specificoligonucleotide primers were designedaccording to the gene sequences of Newcastle disease virus(NDV) and the sequence difference at fusion gene cleavage sites between virulent and avirulent strains.A multiplex RT-PCR method was developed to differentiate virulent NDV strains from avirulentNDV strains.Results showed that a specific band of 442 bp and a common band of 671 bp were amplified from the virulent NDV strains,while a specific band of 252 bp and a common band of 671 bp were amplified from the avirulent NDV strains,and as little as 100 pg of NDV RNA can be detected in a few hours,by the developed multiplex RT-PCR method.