禽流感病毒H5、H9亚型的多重RT-PCR鉴别诊断

Differential diagnosis of H5、H9 subtype of avian influenza virus by multiplex RT-PCR

根据禽流感病毒H5、H9亚型的HA基因序列 ,设计了两对RT_PCR引物 ,同时对H5、H9亚型进行了多重RT_PCR扩增 ,得到了两条清晰的目的带。将目的带回收并进行测序 ,同源性比较结果证明其为禽流感H5、H9亚型的HA基因序列 ,作者对多重RT_PCR进行反应条件优化以及敏感性与特异性测定 ,证明该方法的敏感性和特异性都比较好。初步应用于 30份临床病料 ,其检测结果与病毒的分离培养一致 ,比电镜和琼扩的检出率高 ,与其他NDV、IBV的病料无交叉反应 ,可用于禽流感病毒H5。

This article reports the method of detect H5?H9 subtypes of AIV by Multiplex RT-PCR,which is hiding in poultry and the products of poultry.Two sets of specific primers were designed according to the HA sequences registered in GenBank of different strains of H5?H9 subtypes.Then the optimal conditions of Multipex RT-PCR were determined by orthongonal assay.The rudimentary application of the Multiplex RT-PCR has the same results with Seperate Cultivation of the virus,the positive results acquired is higher then AG and EM.The method described is very practable,sensitive,specific and simple.It could be used in the differential diagnosis of H5?H9 subtypes of AIV.