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牛病毒性腹泻-粘膜病病毒地方株动物回归试验
作者姓名:张宝宁  赵月兰  秦建华  郭红斌  包永占  田席荣
作者单位:河北农业大学动物科技学院 河北保定071000(张宝宁,秦建华,包永占),河北农业大学中兽医学院 河北定州073000(赵月兰,郭红斌),廊坊市职业技术学院 河北廊坊102800(田席荣)
基金项目:国家科技攻关计划“奶牛主要疫病防治技术研究与产业化示范”部分内容(2002BA518A10):河北省畜牧局科技攻关计划”河北省奶牛病毒性腹泻病毒分子变异机制”(2002-4).
摘    要:【目的】自流产奶牛粪便及血液中分离得到的非细胞病变型牛病毒性腹泻-粘膜病病毒(BVDV),为了解分离毒株的特性,进行回归动物试验;【方法】将此分离毒回归2月龄健康犊牛,接毒25天后扑杀剖检,观察临床症状及病理变化。自犊牛体内采取脾、骨髓、肠淋巴结及血液接种MDBK细胞进行培养,分离病毒,进行细胞传代,将此细胞毒进行RT-PCR检测;【结果】犊牛表现为典型的急性病毒性腹泻症状和病变。自病料分离病毒经MDBK细胞盲传至第9代,均未出现细胞病变。将此细胞毒进行RT-PCR检测,扩增出相应的665bp的片段;【结论】将分离株接种易感犊牛以复制出BVD-MD病症,并从试验牛体内分离得到BVDV,从而进一步确证分离到的毒株属BVDV。

关 键 词:牛病毒性腹泻-粘膜病  回归试验  RT-PCR

Regression Test of an Isolated Strain of Bovine Viral Diarrhea Virus
Authors:Zhang Baoning  Zhao Yuelan  Qin Jianhu  Guo Hongbin  Bao Yongzhan  Tian Xirong
Institution:1,College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001; 2,College of Traditional Chinese Veterinary Medicine, Hebei Agricultural University, Dingzhou 073000; 3,3Langfang Vocation-technical Teachers College, Langfang 0650
Abstract:Objective] The non-cytopathic (NCP) biotype strain of bovine viral diarrhea virus was isolated from the dejection and blood of aborted bovine. Regression test was progressed in order to realizing characters of isolated strain. Method] A healthy male calf of two months-old was inoculated against the MDBK culture of the isolated strain. After treatment, clinical symptoms and pathological lesions were observed and the calf was dissected after 25 days. The virus was isolated from spleen, marrow, lymph node and blood of the calf and was propagated blindly in MDBK cells. The isolated virus was identified by RT-PCR. Results] The results indicated that typical symptoms were seen and serial pathological lesions same as acute bovine viral diarrhea appeared in calf. The isolated virus presented non-cytopathic effect (NCPE) in MDBK cell culture for 9 generations. The infected MDBK culture was detected by RT-PCR, and the expected 665bp fragment was obtained. Conclusion] Typical symptom was seen, and bovine viral diarrhea virus was isolated from the calf. The results indicated the isolated strain was bovine viral diarrhea virus.
Keywords:Bovine viral diarrhea virus  Regression text  RT-PCR
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