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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。
The purpose of Yersinia enterocolitica (Yersinia) and brucella are the main pathogen of zoonotic bacterial disease, serological testing, the two often cross-react, especially O: 9 type Ye The bacteria and Brucella interference is more obvious. The purpose of this experiment is to discriminate between the two pathogenic bacteria at the DNA level. Methods Based on the outL gene of Yersinia pestis, a set of primers was designed to detect loop-mediated isothermal amplification (LAMP). The two kinds of bacteria were tested using this specific, sensitive and simple nucleic acid detection technique Detection. Results LAMP method for the detection of Yersinia positive results, the test results of Brucella negative. Conclusion The two pathogenic bacteria were successfully discriminated at the DNA level. And provide applicable primers for the establishment of LAMP detection method for the non-specific interference of Yersinia. Meanwhile, the invention also provides a reference for the development and application of LAMP technology and the design of LAMP primer.