论文部分内容阅读
目的建立一种快速检测霍乱弧菌的方法。方法利用霍乱弧菌的hlyA基因设计特异性引物,建立LAMP检测方法,以及优化反应体系,并进行LAMP的特异性和灵敏度实验,同时与普通PCR和荧光定量PCR进行比较。结果成功建立霍乱弧菌的LAMP检测方法,得到最优的反应体系,在特异性实验中,霍乱弧菌均呈阳性,而副溶血弧菌、溶藻弧菌、最小弧菌、1梅氏弧菌等均为阴性;在灵敏度实验中,霍乱弧菌的最低检测限为6.2pg/tube,是普通PCR的1000倍,比荧光定量PCR低一个数量级。结论LAMP方法具有很高的特异性和灵敏度,可用于霍乱弧菌的检测。
Objective To establish a rapid detection of Vibrio cholerae. Methods Based on the hlyA gene of Vibrio cholerae, specific primers were designed to establish LAMP assay, and the reaction system was optimized. The specificity and sensitivity of LAMP assay were compared with those of PCR and fluorescence quantitative PCR. Results Vibrio cholerae LAMP detection method was successfully established, and the optimal reaction system was obtained. Vibrio cholerae were all positive in the specific experiments, but Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio mimicus, In the sensitivity experiment, the detection limit of Vibrio cholerae was 6.2pg / tube, which was 1000 times of that of normal PCR and one order of magnitude lower than that of fluorescence quantitative PCR. Conclusion The LAMP method has high specificity and sensitivity and can be used for the detection of Vibrio cholerae.