仔猪致泻性大肠杆菌毒力因子多重PCR检测方法的建立与应用  

Establishment of multiplex PCR detection method for diarrhea-causing Escherichia coli virulence factor in piglets in Jilin Province

作　　者：毕斓婷 冀亚路 袭恒豫 张昊 陈椿桦 姜秋杰[2] 付殿国 牛伟 孙长江 冯新 吕伟 顾敬敏 韩文瑜 BI Lanting;JI Yalu;XI Hengyu;ZHANG Hao;CHEN Chunhua;JIANG Qiujie;FU Dianguo;NIU Wei;SUN Changjiang;FENG Xin;LYU Wei;GU Jingmin;HAN Wenyu(College of Veterinary Medicine,Jilin University,Changchun 130062,China;Jilin Province Animal Disease Prevention and Control Center,Changchun 130062,China;College of Animal Science and Technology,Jilin Agricultural Science and Technology University,Jilin City,Jilin 132301,China;Gongzhuling Xinyuan Animal Husbandry Company,Gongzhuling,Jilin 136100)

机构地区：[1]吉林大学动物医学学院,吉林长春130062 [2]吉林省动物疫病预防控制中心,吉林长春130062 [3]吉林农业科技学院动物科技学院,吉林吉林市132301 [4]吉林省公主岭市鑫源木业,吉林公主岭136100

出　　处：《中国兽医学报》2022年第2期250-255,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(U19A2038,32072824,31872505)。

摘　　要：本研究以致泻性大肠杆菌(DEC)的4种毒力因子EAST1、LT、STa和STb为模板设计特异性引物并优化各项反应条件,以建立仔猪DEC的多重PCR检测方法。该方法具有较好的特异性,只对DEC产生特异性条带,对非DEC及其他病菌则呈阴性。利用所建立的多重PCR方法对吉林省各猪场采集的样本进行检测,结果表明,在测定的猪场中,携带EAST1毒力基因的阳性菌株引起的大肠杆菌感染广泛存在,携带STb毒力基因的阳性菌株次之,携带STa及LT毒力基因的阳性菌株引起的大肠杆菌感染相较于以上2个毒力因子在所测猪场中占比略低。综上所述,本方法的建立为DEC中常见毒力因子的快速检测及分子流行病学调查提供了新的手段。In this study, four virulence factors of diarrhea-causing Escherichia coli(DEC),EAST1,LT,STa and STb were used as templates to design specific primers and optimize various reaction conditions to establish a multiplex PCR detection method for piglet DEC. This method has good specificity and only produces specific bands for DEC,but is negative for non-DEC and other pathogens.The established multiplex PCR method was used to detect samples collected from pig farms in Jilin Province.The results showed that E.coli infections caused by positive strains carrying EAST1 virulence genes were widespread in the tested pig farms, and they carried STb virulence genes.Strains were followed by E.coli infections caused by positive strains carrying STa and LT virulence genes.Compared with the above two virulence factors, the proportion of the tested pig farms was slightly lower.In summary, the establishment of this method provides a new method for the rapid detection and molecular epidemiological investigation of common virulence factors in DEC.

关 键 词：致泻性大肠杆菌 毒力因子 多重PCR 

分 类 号：S858.28[农业科学—临床兽医学]