摘要
牛流行热病毒(BFFV)是引发世界范围内牛流行热最主要病原之一,给奶牛业的发展造成严重危害。本研究通过RT-PCR对疑似牛流行热奶牛抗凝血样品进行检测,选取阳性样品颅内接种3日龄乳鼠,7日后乳鼠死亡,鼠脑组织通过PCR扩增出BFFV目的条带,乳鼠颅内重复接种3次,乳鼠发病死亡时间明显提前。选取阳性脑组织接种BHK-21细胞,盲传3代后出现明显细胞病变,特异性引物扩增获得目的条带,测序结果与牛流行热病毒序列一致,表明成功分离到一株牛流行热病毒。
Bovine ephemeral fever virus is one of the most pathogens leading to the bovine ephemeral fever worldwide, which has caused serious harm to the development of dairy industry. In this study, the anticoagulant sample of suspected bovine ephemeral fever from Henan were tested by RT-PCR, then positive samples was inoculated intracerebrally into sucking mice with 3 days of age. After 7 days, the suckling mice died. The target band of BFFV was identified by PCR from rat brain. After repeated intracranial inoculation for 3 times, the death time of sucking mice were significantly advanced. Followed by inoculation of positive samples into BHK-21 monolayer cells, the CPE was obvious after three blind inoculation. The target band were obtained with the specific primers, and the sequencing was consistent with the bovine ephemeral fever sequence. The results showed that a bovine ephemeral fever virus was successfully isolated.
作者
楚会萌
任亚初
程凯慧
蒋仁新
张云飞
解晓莉
张亮
杨宏军
Chu Huimeng;Ren Yachu;Cheng Kaihui;Jiang Renxin;Zhang Yunfei;Xie Xiaoli;Zhang Liang;Yang Hongjun(Dairy Cattle Research Center,Shandong Academy of Agricultural Sciences,Jinan 250131,China)
出处
《山东农业科学》
2019年第3期111-114,共4页
Shandong Agricultural Sciences
基金
"十三五"国家重点研发计划项目(2016YFD0500904
2017YFD0500904)
现代农业(奶牛)产业技术体系科学家岗位项目(CARS-36)
山东省农业科学院农业科技创新工程项目(CXGC2018E14
CXGC2016B14
CXGC2016A10)
