猪呼肠孤病毒SC-A株的分离鉴定及σ2基因的克隆与序列分析

Isolation of reovirus SC-A strain from fecal of diarrhea pigs and sequence analysis of σ2 gene

通过在病毒培养液中添加胰酶的方法,从仔猪腹泻粪样中分离并鉴定了1株能在Vero细胞上稳定产生CPE,并以细胞颗粒增多、肿胀、漂落为特征的猪呼肠孤病毒SC-A株。在感染细胞中,病毒胞浆包涵体及特征性微管样结构明显,病毒粒子多呈典型的晶格状排列,大小约70 nm。S2基因的克隆测序结果表明,SC-A S2全基因(DQ396805)大小为1 331 bp,其开放阅读框编码一个由418个氨基酸组成,相对分子量约为47 140的σ2蛋白;与标准株T1L、T2J、T3D的核苷酸序列和推导氨基酸序列的同源性分别为86.9%、76.7%、85.4%及94.6%、93.3%、98.3%。以σ2氨基酸序列构建的进化树分析显示,标准株及野外分离株可被划分为A(T3)、B(T2)、C(T1)3个群;除T3C31外,其余3型野外分离株及SC-A株和所有血清1型均位于C群中。

Using Vero cell line and by the method of adding the trypsin into the viral culture MEM,reovirus SC-A strain was isolated form the pigs diarrhea fecal.The stable CPE characters of the reovirus,including cell granulations increasing,cellular swelling and shedding were observed.In the cytoplasm of infected cell,the virion of diameter approximately 70 nm,shows typical lattice arrangement,the cytoplasmic inclusion bodies and the distinctive microbule-like structure.The S2 gene(DQ396805)of reovirus SC-A,obtaine...