西尼罗病毒的RT-PCR检测与鉴定

建立西尼罗病毒敏感、特异、快速的RT-PCR检测方法用于实验室诊断和流行病学监测。采用一步RT-PCR和套式PCR法对西尼罗病毒感染的乳鼠脑和细胞培养上清进行扩增，并对扩增产物进行序列测定。两种方法均可分别从两种组织中扩增出与预期大小相一致的片段，套式PCR法比一步RT-PCR法更为敏感，该扩增片段与西尼罗病毒埃及Eg101株相应序列的同源性为99％。

Detection and identification of West Nile virus by RT-PCR

In order to establish a sensitive, specific and rapid RT PCR assay for laboratory diagnosis and epidemiology surveillance of West Nile virus,the cDNA fragments of West Nile virus were amplified from the brains of infected suckling mice and the supernatants of infected cell culture by one step RT PCR and nested PCR assays, then the amplified fragment was sequenced. The sizes of the amplified fragments from two infected tissues was demonstrated by two PCR assays to be equal to those of the expected products.The nested PCR assay was more sensitive than RT PCR assay.The homology between the amplified fragment of the Chin 01 strain and Eg101 strain was 99%.