摘要
牛病毒性腹泻病毒（Bovine viral diarrhoea virus,BVDV）不同的基因型在抗原性方面存在差异,使疫苗免疫难以收到预期效果。本试验对分离到的BVDV石河子株和其他石河子分离株进行基因型鉴定,通过对病毒的遗传分类研究,为该病的防治和开发新的基因工程疫苗提供一定依据。采用免疫学试验、电镜观察、理化特性测定以及PCR技术、同源性比较等方法分离、鉴定1株BVDV石河子分离株,命名为BVDV shz132株,并应用系统发生分析方法鉴定该株与其他石河子分离株Manasis、hihezi148和Letuyi株的基因型。将免疫学鉴定为阳性的样品进行病毒分离,电镜观察分离的BVDV shz132株病毒直径为40~60 nm,为有囊膜的球形颗粒。理化鉴定结果与参考株BVDV Oregon C24V一致。根据BVDV基因组5-′UTR基因设计引物在发病牛淋巴结、肠道分泌物制备的悬液及其分离的病毒细胞培养物中均扩增得到了大小为267 bp的片段。核苷酸同源性分析结果表明BVDV shz132株与BVDV Osloss株的同源性最高（97.5%）,BVDV Manasi株等与BVDV NADL株的同源性最高（97.8%）;系统发生分析表明上述所有石河子分离株均属于BVDV-Ⅰ型。
Bovine viral diarrhoea virus（BVDV） has cross-infection with classical swine fever virus（CSFV） and border disease virus of sheep（BDV）.Furthermore,different genotypes of BVDV potentially led to the failure of vaccination.Genotype BVDV is important to BVD prevention and new vaccine development.A BVDV strain originated from Shihezi area in Xinjiang autonomous district,namely BVDV shz132,was isolated and identified by immunological method,electron microscope observation,physico-chemical property analysis,PCR detection and homology comparison.The phylogenetically analyses was carried out including other isolates named as Manasi,shihezi148 and Letuyi also originated from Shihezi area in Xinjiang autonomous district.Viral particle with the envelope was observed to be round and about 40-60 nm in diameter.A 267 bp fragment was amplified by reverse transcription polymerase chain reaction（RT-PCR） using lymph node,intestinal tract secreta or cell culture extracts as the templates.The primers were designed based on the 5′UTR gene sequences.The results showed that BVDV shz132 strain had the highest identity（97.5%）with BVDV Osloss strain in homology,while BVDV Manasi,shihezi148 and Letuyi had 97.8% homology with BVDV NADL strain,and all of the isolates originated from Shihezi area in Xinjiang autonomous district mentioned above fell into the BVDV-Ⅰgroup.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2010年第4期433-439,共7页
Chinese Journal of Veterinary Science
基金
国际科技合作项目(2006DFA33740)
