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

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. arc both high infectious and zoonotic bacteria. False-positive scrological reactions due to infections with brucellosis arc problems in tests for Yersinia enterocolitica especially scrotypc O: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 charactcrisitcs of high specificity, sensitivity, easy to operate and simple to get the result was carried out to distinguish Yersinia enterocolitica from Brucella with the primers that were designed by oulL gene from Yersinia enterocolitica. Results The result of Yersinia 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 spccific primers to establish a LAMP method for detection of Yersinia enlerocolilica and for giving some suggestions to design LAMP primers for the development and application of LAMP method.