西尼罗热病毒RT-PCR检测方法的建立

Construction of RT-PCR for Detection of West Nile Virus

参考Genebank发表的西尼罗热病毒(WestNilevirus,WNV)E糖蛋白基因序列,自行设计合成一对引物,对WNV进行RT-PCR扩增,产物经琼脂糖电泳分析,呈现一条约400bp的条带,将其克隆入pMD18-T-Vector载体中,并进行序列测定,与已发表的WNV基因比较发现,核苷酸的同源性为99.7%,证实为WNV的E基因,通过对样品多次检测,都能扩增出一条约400bp的条带,表明该方法比较稳定。

According to the Genebank published sequence of west nile virus ,a pair of primers were designed and synthesized in order to amplify E gene of West Nile virus.The E gene was obtained by reverse transcript polymerase chain reaction(RT-PCR) ,subsequently cloned into pMD18-T-Vector.The nucleotide of West Nile virus E gene was compared with the corresponding sequences of published sequence of WNV,the nucleotide homology was 99.7% suggesting,that it is E gene of West Nile virus..