禽流感病毒和新城疫病毒二联RT-PCR检测方法的建立

参照国内外已发表的禽流感病毒(AIV)和新城疫病毒(NDV)的基因序列及其相关的RT-PCR检测方法,根据禽流感病毒M蛋白基因和新城疫病毒NP基因各设计一套特异性通用引物,扩增目的带分别为600bp和340 bp。通过对相关病毒检测,建立了AIV和NDV通用型二联RT-PCR检测方法。该方法具有快速、敏感、特异等优点,可为AIV和NDV的检测、流行病学调查及疫苗使用等奠定基础。

Establishment of AIV and NDV by Multiplex RT-PCR

According to the published gene sequence of Avian influenza virus(AIV) and Newcastle disease virus(NDV) and the detecting methods of RT-PCR reported,matrix(M) gene of AIV and nucleoprotein(NP)gene of NDV were considered as the target gene sequences of multiple RT-PCR.A set of universe primers were designed for detecting AIV and NDV,respectively in the conserved M gene of AIV and NP gene of NDV.The products of RT-PCR is 600 bp and 340 bp respectively.The optimal condition of multiple RT-PCR for detection of AIV and NDV by RT-PCR was determined through orthogonal assay.The specificity of the multiple primers were examined by RT-PCR using template extracted from other avian virus.The sensitivity of RT-PCR were also determined by a seral dilution of RNA from AIV and NDV.This study have paved the way for testing AIV and NDV,defining pathogenic potential,using vaccines against AIV and NDV and so on.