日本乙型脑炎病毒的分离与鉴定

收集一例疑为乙型脑炎病毒感染的种公猪肿大的睾丸病料,并将其处理制成匀浆过滤后,用乳鼠脑内接毒和细胞接毒相结合的方法盲传并分离病毒,然后设计一对PrM/E基因的特异性引物,采用RT-PCR方法对所分离的病毒进行鉴定,并将其PrM/E基因扩增产物测序,测序结果与乙型脑炎GenBank登陆的SA14-14-2株(AF315119)和SA14株(U14163)进行比较。结果表明,所分离病毒的PrM/E基因序列与JEV强毒株SA14和弱毒株SA14-14-2相应序列同源性分别为98.1%和97.1%,证实分离毒株为乙型脑炎病毒。

Isolation and identification of Japanese encephalitis virus

The swelling testicles sample from a boar suspected of Japanese encephalitis virus (JEV) infection was collected, made into homogenate and filtrated. The filtrate was injected into rat brain and passaged on cell monolayer to obtain the viral strain. A pair of PrM/E gene-specific primers was designed and the viral strain was identified by RT-PCR method. Then the PCR product was further sequenced. The obtained PrM/E nucleotide sequence was compared with those of SA14-14-2 and SA14 strains available in GenBank. The result showed that the PrM/E gene sequence shared 98. 1% and 97.1% of nucleotide homology with SA14 and SA14-14-2 strains, respectively. The result proved that the isolated virus was JEV.