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通过比较现有几种可视化检测环介导等温扩增反应产物的方法,建立一种可视化检测变形杆菌的方法。利用已有的LAMP检测变形杆菌的方法作为出发检测体系,比较了焦磷酸镁反应沉淀观察法和分别单独添加核酸染料SYBR Green I、孔雀绿(Malachite Green)、钙黄绿素-氯化锰(Calcein-MnCl2)的可视化检测方法,结果显示,焦磷酸镁沉淀的指示效果较差,SYBR Green I法会造成实验场所的交叉污染,孔雀绿法指示反应不够灵敏,钙黄绿素-氯化锰法综合考虑较优;并证明该方法用于可视化LAMP检测变形杆菌,具有良好的特异性,灵敏度达到4.8CFU/mL。试验证明钙黄绿素-氯化锰法是一种较优的LAMP可视化检测法。
By comparing the existing methods for visualizing the ring-mediated isothermal amplification reaction products, a visualization method for detecting Proteus was established. The method of detecting Lactobacillus by LAMP was used as a starting detection system, magnesium pyrophosphate reaction precipitation observation method and the method of separately adding the nucleic acid dyes SYBR Green I, Malachite Green, Calcein- MnCl2). The results showed that the indication of magnesium pyrophosphate precipitation was poor, and the SYBR Green I method could cause cross-contamination in the experimental sites. The malachite green method indicated that the reaction was not sensitive enough, and the calcein-manganese chloride method considered The method has been proved to be good for the visualization of LAMP detection of Proteus, with a sensitivity of 4.8 CFU / mL. Experiments show that calcein - manganese chloride method is a better visualization of LAMP detection method.