仔猪腹泻粪样中猪呼肠孤病毒的分离鉴定  

Isolation,Identification of Porcine Reovirus from Diarrhea Feces of Pigs

作　　者：曾智勇[1] 郭万柱[1] 徐志文[1] 宋振辉[1] 殷华平[1] 王新[1] 王小玉[1] 

机构地区：[1]四川农业大学动物生物技术中心

出　　处：《畜牧兽医学报》2007年第6期574-580,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金："教育部长江学者和创新团队发展计划"项目(IRTO555)

摘　　要：通过在病毒培养液中添加胰酶的方法,笔者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,即以细胞颗粒增多、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,直径约70 nm。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1个由418个氨基酸残基组成、相对分子量约为47.14 ku的σ2蛋白;与标准株T1L、T2J、T3D的核苷酸序列相似性分别为86.9%、76.7%、85.4%,推导氨基酸序列相似性分别为94.6%、93.3%、98.3%。在以σ2氨基酸序列构建的进化树中,标准株及野外分离株可被划分为A、B、C 3个大的进化分支;除T3C31外,其余3型野外分离株及SC-A株和所有血清1型病毒株均位于进化分支C中。Using Vero cell line and adding the trypsin into the viral culture MEM, reovirus SC-A strain was isolated form the pigs＇ diarrhea feces. The stable CPE characters of the reovirus were as follows： cell granulations increasing, cellular swelling and shedding. The virions of approximately 70 nm diameter, which shows typical lattice arrangement in cytoplasm of infected cells and the cytoplasmic inclusion bodies and the distinctive microbule-like structure occured. The S2 gene （DQ396805）of reovirus SC-A, obtained by RT-PCR, was 1 331 bp in length and encodes the a2protein, which with relative molecular weight of 47.14 ku. Sequence analysis of nucleotides indicated that the reoviurs SC-A S2 gene shared 86.9%, 76.7% and 85.4% similarity with standard strains TIL, T2J and T3D, respectively. And the similarity of deduced amino acid sequence of reoviurs SC-A S2 gene were 94. 6%, 93. 3% and 98. 3% with standard strains T1L, T2J and T3D, respectively. Phylogenetic trees based on a2 shows standard strains T1L, T2J and T3D were divided into branch A, B and C, respectively. Except T3C31 field isolated strain, serotype 3 of the other field isolate strains, SA-C strain, all of serotype 1 strains were located in branch C of phylogenetic tree.

关 键 词：猪 呼肠孤病毒 分离 鉴定 

分 类 号：S852.659.4[农业科学—基础兽医学]