变形杆菌环介导等温扩增检测法的可视化方法的比较

发布时间：2018-03-11 11:44

本文选题：环介导等温扩增　切入点：可视化检测　出处：《河北农业大学学报》2016年03期 　论文类型：期刊论文

【摘要】：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。
[Abstract]:By comparing several methods of visual detection of isothermal amplification products mediated by ring, a visual detection method for Proteus was established. The existing LAMP method for detection of Proteus was used as the starting detection system. The visual detection methods of magnesium pyrophosphate reaction precipitation observation method and separately added nucleic acid dye SYBR Green I, malachite Greenfen, Calcein-MnCl2) were compared. The indicating effect of magnesium pyrophosphate precipitation is poor. The method of SYBR Green I will cause cross contamination of the experimental site, the indicator reaction of malachite green method is not sensitive enough, and the comprehensive consideration of calcium xanthophyllin and manganese chloride method is better. It is proved that this method has good specificity and sensitivity of 4.8 CFU / mL for visual detection of Proteus by LAMP. It is proved that calcium xanthophyllin-manganese chloride method is an excellent visual detection method for LAMP.
【作者单位】：河北农业大学食品科技学院/河北省农产品加工工程技术中心;河北农业大学理工学院;
【基金】：国家自然科学基金资助项目(31371772和31071494) 河北省科技计划项目(152776120D)
【分类号】：R378.2

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