新疆部分地区牛病毒性腹泻病的流行病学调查及其病原的分离与鉴定

发布时间：2018-10-16 19:47

【摘要】：牛病毒性腹泻/粘膜病毒(Bovine viral diarrhea/mucosal disease virus,BVD/MDV)是黄病毒科,瘟病毒属的成员,具有极其严重的危害性。可引起一种呈世界性分布的疾病—牛病毒性腹泻/粘膜病,给畜牧业造成了极大的危害。随着畜牧业的快速发展,其危害性也日益严重。目前,该病尚未有有效的防控措施。因此,获得牛病毒性腹泻的流行病学资料,为该病的后续防控提供充足的理论基础。本研究采用血清学方法和分子生物学方法对新疆16个地区不同牛场采集的血清、全血和粪便样品进行检测,并对检测到的阳性样品进行病原学分离鉴定。通过对1171份样品进行血清学检测,其中抗体阳性率为81.9%;抗原阳性率为1.5%。应用RT-PCR方法对566份样品进行检测,从中检测到了21株阳性BVDV样品。对测序结果进行序列同源性比对分析,绘制其进化树,并初步绘制出了新疆部分地区BVDV病毒的地理生态分布图,其中17株为BVDV I型,4株为BVDV II型。应用细胞培养分离病毒,利用RT-PCR方法和直接荧光免疫检测进一步鉴定病毒,最后测定病毒的病毒滴度和对阳性血清的中和效价,并进行病毒的理化特性和生物学特性分析。结果发现7株为致细胞病变型,14株为非致细胞病变型。所测得的病毒滴度为1.04×103~6.02×108 TCID50/0.1mL,21株阳性样品对BVDV阳性血清的中和效价为1:21~1:25。分离的病毒对温度、乙醚、氯仿和胰酶很敏感。新疆地区的BVDV流行株有BVDV 1b亚型、I型未知亚型和BVDV II型。研究成果为BVDV的预防和控制提供了理论基础和参考依据。
[Abstract]:Bovine viral diarrhea / mucosal virus (Bovine viral diarrhea/mucosal disease virus,BVD/MDV) is a member of the family Flavoviridae. It can cause a worldwide disease-bovine viral diarrhea / mucosal disease, which has caused great harm to animal husbandry. With the rapid development of animal husbandry, its harmfulness is becoming more and more serious. At present, there is no effective prevention and control measures. Therefore, to obtain the epidemiological data of bovine viral diarrhea provides sufficient theoretical basis for the follow-up prevention and control of the disease. In this study, serological and molecular biological methods were used to detect serum, whole blood and fecal samples collected from 16 cattle farms in Xinjiang, and the positive samples were isolated and identified by etiology. By serological examination of 1171 samples, the positive rate of antibody and antigen were 81.9 and 1.5 respectively. RT-PCR method was used to detect 566 samples, from which 21 positive BVDV samples were detected. The sequence homology analysis was carried out, the evolutionary tree was drawn, and the geographic and ecological distribution of BVDV virus in some regions of Xinjiang was preliminarily plotted. Among them, 17 strains were BVDV I type and 4 were BVDV II type. The virus was isolated by cell culture. The virus was further identified by RT-PCR method and direct fluorescence immunoassay. Finally, the virus titer and neutralization titer to positive serum were determined, and the physicochemical and biological characteristics of the virus were analyzed. The results showed that 7 strains were cytopathic and 14 were non-cytopathic. The titer of the virus was 1.04 脳 10 ~ 3 and 6.02 脳 10 ~ 8 TCID50/0.1mL,21 strain. The neutralization titer of BVDV positive serum was 1: 21 1: 25. The isolated virus is sensitive to temperature, ether, chloroform and trypsin. The research results provide theoretical basis and reference for prevention and control of BVDV.
【学位授予单位】：新疆农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.23

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