可视环介导恒温扩增技术检测实验猴小肠结肠炎耶尔森菌

Visible Loop-mediated isothermal amplificationmethod for examination of experimental monkey Yersinia enterocolitica

本研究在传统环介导恒温扩增反应体系中引入羟基萘酚蓝指示剂,对反应体系进行高温-低温前处理和添加甜菜碱等优化措施提高试验敏感性,液面覆盖矿物油降低气溶胶污染,据此建立的猴小肠结肠炎耶尔森菌可视LAMP检测方法结果判定直观明了,敏感度为6 CFU/反应,无非异性。

In this study,the indicator hydroxynaphthol blue was added to reaction system of the traditional Loop-mediated isothermal amplification. The reaction system was prepared by heating-cooling and betaine was added to the reaction system to improve the sensibility. The liquid surface was covered by mineral oil. Base on these,we built a visible loop-mediated amplification method for detecting experimental monkey Yersinia enterocolitica. The visible LAMP/s sensibility was 6 cfu/reaction. The visible LAMP showed intuitive and specific results,and the method was more efficient and had higher detection rate than germiculture.