PCR法检测大肠杆菌的增殖条件优化研究  

Optimization of pre-enrichment protocol of PCR-based method for detection of Escherichia coli

作　　者：吕晓萌[1,2] 胡文忠[2] 冯叙桥[1,3] 何煜波[2] 董妍[4] 

机构地区：[1]沈阳农业大学食品学院,辽宁沈阳110866 [2]大连民族学院生命科学学院,辽宁大连116600 [3]渤海大学食品科学研究院,辽宁省食品安全重点实验室,技术研究中心,辽宁锦州121013 [4]大连工业大学食品学院,辽宁大连116034

出　　处：《食品工业科技》2014年第17期134-136,146,共4页Science and Technology of Food Industry

基　　金：国家自然科学基金项目(31172009);国家科技支撑计划项目(2012BAD38B05);大连科技计划项目(2012E13SF106)

摘　　要：目的:比较不同培养模式与不同添加剂对大肠杆菌增殖效果的影响。方法:本实验以大肠杆菌为目标菌,研究了2种培养模式(振荡与静止培养)、2种添加剂(甘露醇与乳糖)对大肠杆菌增殖效果的影响,并采用PCR法检验其增殖效果。结果得出:振荡培养的增殖效果优于静止培养。在LB培养基中添加甘露醇和乳糖可起增殖作用,其中0.3g/100mL甘露醇与1.0%乳糖复配4h后,大肠杆菌的增殖效果最佳。检测中使用0.3g/100mL的甘露醇和1.0%的乳糖,并辅以37℃、150r/min的振荡培养即可获得良好效果。结论:本方法在6h内即可完成大肠杆菌的增殖及PCR检测,可用于食品中大肠杆菌的快速检测。Objective.To compare the enrichment effect of different culture methods and different medium additives on Escherichia coil Method; This study was to investigate the enrichment effect of 2 culture methods （ static cultivation and shaking cultivation） and 2 medium additives （ lactose and mannitol） on Escherichia coli, and to evaluate the enrichment effect by PCR.Results.The results indicated that shaking cultivation was better than static cultivation.The effect of LB medium adding mannitol and lactose was beneficial to the growth of Escherichia coil When the dosage of lactose and mannitol were 1.0% and 0.3g/100mL, the effect of enrichment was best. The enrichment time was 4h.The detection could get good results by using 1.0% lactose and 0.3g/100mL mannitol with shaking cultivation at 37℃.Conclusion ：The enrichment and PCR detection of Escherichia coil can be accomplished in 6h,which can be used for rapid detection of Escherichia coil in foods.

关 键 词：PCR检测 大肠杆菌 前增殖 

分 类 号：TS255.3[轻工技术与工程—农产品加工及贮藏工程]