区别检测小肠结肠炎耶尔森氏菌与布鲁氏菌LAMP引物的设计与分析  

Design and analysis of LAMP primers used in differentially detecting Yersinia enterocolitica and Brucella spp

作　　者：徐云明[1] 邹德颖[1] 唐峰[1] 郭兴[1] 王光明[1] 宋杰[1] 刘楠楠[1] 周玉[1] 柳增善[1] 卢士英[1] 李兆辉[1] 任洪林[1] 

机构地区：[1]人兽共患病研究教育部重点实验室吉林大学人兽共患病研究所畜牧兽医学院,吉林长春130062

出　　处：《中国实验诊断学》2013年第1期19-22,共4页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金项目(30901070);吉林大学基本科研业务费项目资助(421060516206)

摘　　要：目的小肠结肠炎耶尔森氏菌(耶氏菌)和布鲁氏菌都是人兽共患细菌病的主要病原菌,血清学检测过程中,两者常出现交叉反应,尤其是O:9型耶氏菌与布鲁氏菌干扰更为明显。本实验目的是在DNA水平上区别检测这两种致病菌。方法根据耶氏菌outL基因设计一套可用于环介导等温扩增(LAMP)检测方法的引物,拟采用该项特异性强、灵敏性高、操作简单的核酸检测技术,对两种菌进行检测。结果 LAMP方法检测耶氏菌结果为阳性,检测布鲁氏菌结果为阴性。结论成功地在DNA水平上区别检测这两种致病菌。并且为建立耶氏菌的非特异性干扰小的LAMP检测方法提供可应用的引物,同时也为LAMP技术的开发应用以及LAMP引物设计提供应用参考。Objective Yersina enterocolitica and Brucella spp. are both high infectious and zoonotic bacteria. False- positive serological reactions due to infections with brucellosis are problems in tests for Yersinia enterocolitica especial- ly serotype 0：9. In this study, Loop-mediated isothermal amplification （LAMP） was used to distinguish the Yersinia enterocolitica from the Brucella at the molecular level. Methods LAMP which has the characterisitcs of high specifici- ty,sensitivity,easy to operate and simple to get the result was carried out to distinguish Yersinia enteroeolitica from Brucella with the primers that were designed by outL gene from Yersinia enterocolitica. Results The result of Yersin- ia enterocolitica was positive and the one of Brucella was negative. Conclusion This study distinguishes the Yersinia enterocolitica from the Brucella successfully at the molecular level at the same time is beneficial for offering high specific primers to establish a LAMP method for detection of Yersinia enterocolitica and for giving some suggestions to de- sign LAMP primers for the development and application of LAMP method.

关 键 词：小肠结肠炎耶尔森氏菌 布鲁氏菌 环介导等温扩增 

分 类 号：R516.7[医药卫生—内科学]