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目的建立并优化食品及环境样品中肠球菌的快速检验方法。方法用mEI培养基作为选择性培养基,肠球菌tuf基因为靶基因设计引物,建立食品及环境样品中肠球菌的快速检验方法。通过添加肠球菌和非肠球菌标准菌株,验证方法的检测限、灵敏度及特异性,并使用优化后的方法对北京市丰台区某大型生鲜猪肉集贸市场的地面、墙壁、污水和生鲜猪肉等样品中的肠球菌进行检验。结果所建方法的检测限为1 CFU/100cm2和1 CFU/25 g(mL),且与10种非肠球菌无交叉反应;地面涂抹、墙壁涂抹、污水和生鲜猪肉等样品中均检验出肠球菌,检出率为100%。结论建立了简便、快速、灵敏、特异的肠球菌检验方法;集贸市场内部环境和生鲜猪肉存在肠球菌污染。应加强市售生鲜肉中肠球菌的监测,并采取相应措施降低其污染水平。
Objective To establish and optimize a rapid test method for enterococci in food and environmental samples. Methods Using mEI medium as selective medium and Enterococcus tuf gene as target gene to design primers, a rapid test method of enterococci in food and environmental samples was established. The limits of detection, sensitivity and specificity of the method were validated by adding enterococci and non-enterococcal standard strains. The optimized methods were used to analyze the surface, wall, sewage and freshness of a large-scale fresh pork market in Fengtai District of Beijing Enterococcus in samples such as pork is tested. Results The limits of detection of the proposed method were 1 CFU / 100 cm 2 and 1 CFU / 25 g (mL), and there was no cross-reaction with 10 non-enterococci. The results of the tests on floors, walls, sewage and fresh pork Enterococcus, the detection rate was 100%. Conclusions A simple, rapid, sensitive and specific method for Enterococcus faecalis was established. Enterococci contamination was found in the internal environment of bazaars and fresh pork. Should be strengthened in the market fresh meat in the detection of Enterococcus, and take appropriate measures to reduce its level of pollution.