多重荧光LAMP法在食源性致病菌风险监测中的应用

Application ofmultiple fluorescent LAMP reaction for foodborne pathogenic bacteria detection in risk monitoring

为解决食源性致病菌传统检测方法耗时长、工作量大问题,利用金黄色葡萄球菌femA序列和沙门氏菌invA序列设计了两套LAMP引物,添加荧光染料建立混合体系,通过恒温荧光检测仪对目标DNA进行扩增得到相关曲线,同时对扩增产物进行电泳分析,并且对100批次食品样品进行了效果验证。试验结果表明,该检测方法的普筛性与特异性良好,灵敏度达到71 CFU/mL,检测过程不易污染、耗时短。该方法可应用于部分食源性致病菌的快速初筛。

The problemof traditional pathogenic bacteria detection method is time consuming and the large workload.The objective of the article is to solve the difficulty. Staphylococcus aureus and Salmonella were designed two sets of LAMP primers by invAand femAsequences. The fluorescent was added to the hybrid system. The target DNA getted the amplification and the related curve through the constant temperature fluorescence detector. At the same time, the amplification products was analyzed through the electrophoresis apparatus,and 100 batches of food samples were tested in the end. The method has good screen and specificity. The sensitivity is 71 CFU/mL. The pollution and lowefficiency is hard to happen in the process. The method can be applied to the beginning of the rapid screening of some food-borne pathogens.