广东地区鸭疫里默氏杆菌的分离鉴定及双重PCR检测方法的建立  

Isolation,identification and development of duplex PCR method for detection of Riemerella anatipestifer from Guangdong province

作　　者：魏秀丽[1] 张兴晓[1,2] 姜世金[1] 孙亚妮[1] 刘美丽 杨灵芝 

机构地区：[1]山东农业大学动物医学院,山东泰安271018 [2]山东博莱威生物技术研究所,山东滨州256618

出　　处：《中国兽医学报》2009年第6期721-725,共5页Chinese Journal of Veterinary Science

基　　金：山东省科技攻关项目(2005GG440001);山东省自然科学基金项目(Q2006D04)

摘　　要：对来自广东地区疑似鸭疫里默氏杆菌病的病鸭组织进行了病原菌的分离培养和生化鉴定,确定得到20株鸭疫里默氏杆菌,动物回归试验结果表明,分离的鸭疫里默氏杆菌具有很强的致病性。对这20株鸭疫里默氏杆菌进行的药敏试验表明,分离株已广泛产生耐药性。参照已经发表的2对引物以细菌全菌体为模板建立双重PCR方法,结果均能扩增出2条目的片段,经测序证实为鸭疫里默氏杆菌,而对鸭源大肠杆菌、鸭源禽多杀性巴氏杆菌、鸭源沙门菌和鸭源葡萄球菌的扩增结果均为阴性。说明建立的双重PCR方法能快速、准确的检测出鸭疫里默氏杆菌,并具有高度特异性,可用于鸭疫里默氏杆菌的快速鉴定和快速诊断。Twenty strains of Riemerella anatipestifer （RA） were isolated and identified based on biochemical tests from suspected sick ducks with infectious serositis in Guangdong province and the results of animal experiment showed that the pathogenicity of the isolated RA strains was very high. The result of drug sensitivity test showed twenty RA strains had widely drug-resistance. According to two pairs of published primers and using whole bacteria cells as the template,the duplex PCR was established. The results showed that two fragments were amplified from all detected RA strains, while Escherichia coli , Pasteurella multocida , Salmonella and Staphylococcus aureus strains isolated from duck were negative results. Thus,the duplex PCR assay can he used to rapid identification and diagnosis of RA （infection）.

关 键 词：鸭疫里默氏杆菌 生化鉴定 药敏试验 动物试验 双重PCR 

分 类 号：S852.61[农业科学—基础兽医学]