生鲜牛乳中产肠毒素大肠埃希菌环介导等温扩增技术的建立与应用

Establishment and Application of Enterotoxigenic E. coli Loop-mediated Isothermal Amplification Method from Raw Milk

为了快速检测生鲜牛乳中的产肠毒素大肠埃希菌(enterotoxigenic E.coli,ETEC),防止食源性产肠毒素大肠埃希菌给人类造成的危害,建立快速检测不耐热型产肠毒素大肠埃希菌的环介导等温扩增方法(LAMP)。敏感性和特异性试验结果表明,该方法敏感性高,特异性强,能够从生鲜牛乳中检测到1pg产肠毒素大肠埃希菌DNA,与其他型大肠埃希菌及细菌不发生交叉反应。利用建立的LAMP技术对来自西宁市奶牛场及自由市场的21份生鲜牛乳样品进行检测,6份样品呈阳性,阳性检出率为28.6%,从分子水平为生鲜牛乳中产肠毒素大肠埃希菌的检测提供了一种快速、准确的手段,对保障生鲜牛乳的食品安全有重要意义。

In order to rapidly detect enterotoxgenic Escherichia coli (ETEC) in raw milk and prevent humans being harmed from food borne ETEC,a loop-mediated isothermal amplification (LAMP) method was established for rapid detection of heat-labile ETEC.The results showed that the method was highly sensitive and specific.The detect limit is 1 pg of ETEC DNA from the raw milk,and without cross reaction with other types of E.coli and other bacteria.21 fresh milk samples from dairy cow farms and free markets were tested using the method.6 samples were positive.The positive rate was 28.6%.The results provided the molecular method for detection of ETEC in raw milk.And it is critical important for ensuring the food safety of the raw milk.