摘要
为进一步了解甘肃省某奶牛场牛副结核病的感染情况,采集该场227头奶牛血清进行ELISA抗体检测,结果,检出13头阳性奶牛,阳性率为5.73%。在13头抗体阳性奶牛中随机采集6头表现拉稀症状奶牛的直肠粪便,提取DNA进行荧光定量PCR检测,结果显示,6份粪便样品均为阳性。6份粪便样品用十六烷基氯化吡啶(HPC)处理后接种自行配制的Herrold’s卵黄琼脂培养基,经过7~14周培养得到5株分离菌株。采用IS900特异性序列PCR鉴定和限制性内切酶分析(REA)鉴定,结果表明,5株分离菌株均为副结核分枝杆菌。采用IS1311特异性序列PCR-REA鉴定,结果表明,5株分离菌株均为C型MAP。
In order to understand the infection situation of bovine paratuberculosis in a dairy cattle farm in Gansu Province,paratuberculosis was detected from 227 cattles in this farm by ELISA.A total of 13(5.73%)cattles were positive for Map antibody from 227 sera tested.Feces of 6 cattles with diarrhea were collected randomly from 13 cows with positive for Map antibody,and detected of 6 feces samples were positive by fluorescent quantitative PCR.The fecal samples were treated with hexadecylpyridinium chloride(HPC)and inoculate on Herrold’s Egg Yolk Agar.Five isolates were obtained after7—14 weeks of culture.The isolated strains were verified by restriction endonuclease analysis(REA)of the IS900 specific genome fragment of the MAP.The IS1311 PCR-REA was successfully isolated five Map strains,which both were Map-C.
作者
常塔娜
田莉莉
许芳
樊晓旭
孙淑芳
范伟兴
CHANG Ta-na;TIAN Li-li;XU Fang;FAN Xiao-xu;SUN Shu-fang;FAN Wei-xing(China Animal Health and Epidemiology Center,Qingdao 266114,China;College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2020年第7期874-879,共6页
Chinese Veterinary Science
基金
现代农业(奶牛)产业技术体系建设专项资金(CARS-36)。
关键词
副结核病
副结核分枝杆菌
分离鉴定
paratuberculosis
Mycobacterium avium subsp.paratuberculosis
isolation and identification
