犊牛腹泻粪样中牛呼肠孤病毒的分离鉴定  

Isolation and identification of bovine reovirus from feces of the calves suffered from diarrhea

作　　者：常继涛[1] 李鑫[1] 张亚科[1] 于力[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2008年第9期711-715,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技支撑计划课题(2006BAD04A09)

摘　　要：通过在培养液中添加胰蛋白酶的方法,用MA104细胞从犊牛腹泻粪样中分离并鉴定了1株能稳定产生细胞病变(CPE)的牛呼肠孤病毒,命名为B-19株。RNA聚丙烯酰胺凝胶(PAGE)电泳可见呼肠孤病毒典型的10条核酸节段,呈3∶3∶4排列。电镜检测结果显示,病毒粒子呈典型呼肠孤病毒粒子形态,直径约70nm;病毒L1基因保守区段RT-PCR检测及序列分析表明,扩增出的440bp目的片段符合预期大小,而且其核苷酸序列与GenBank参考毒株L1基因序列具有较高同源性;S1基因序列分析表明,B-19株属于呼肠孤病毒血清1型(ST1)。A bovine reovirus, designated as the B-19 strain, was isolated from the feces of the calves suffered from diarrhea by cell culture in MA104 cell line. This cell culture-adapted strain could produce stable cytopathic effect （CPE） in vitro. The virus has an RNA polyacrylamide gels （PAGE） electrophoretic migration pattern similar to those of the three serotypes of reovirus, with ten nucleotide segments in a 3：3：4 arrangement. The virus particles were of typical reovirus under electronmicroscope, with a diameter of 70 nm. The 440 bp segment of conservative part of L1 gene was amplified by RT-PCR and sequenced, and the DNA segment shared high similarity with that of standard reovirus strains. The B-19 strain was characterized as reovirus serotypel （ST1） by S1 gene sequence analysis.

关 键 词：牛呼肠孤病毒 分离 鉴定 

分 类 号：S852.659.4[农业科学—基础兽医学]