文献类型：期刊文献

中文题名：基于环介导等温扩增技术快速检测阪崎肠杆菌

英文题名：Rapid Detection of Enterobacter Sakazakii Based on Loop-Mediated Isothermal Amplification

作者：岳紫馨[1];梁玉林[2];徐文文[2];刘秀[2];李祖明[1];尹建军[2]

第一作者：岳紫馨

机构：[1]北京联合大学生物化学工程学院,北京100023;[2]中国食品发酵工业研究院有限公司,北京100015

第一机构：北京联合大学生物化学工程学院

年份：2020

卷号：45

期号：6

起止页码：366-371

中文期刊名：食品科技

外文期刊名：Food Science and Technology

收录：北大核心:【北大核心2017】；

基金：北京高等学校高水平人才交叉培养“实培计划”科研类项目。

语种：中文

中文关键词：等温扩增;阪崎肠杆菌;乳粉;基因;检测

外文关键词：isothermal amplification;Enterobacter sakazakii;milk powder;genes;detection

摘要：为实现婴幼儿配方乳粉中阪崎肠杆菌快速检测,建立等温荧光扩增方法特异性检测阪崎肠杆菌。针对阪崎肠杆菌特异性保守ompA基因设计3套引物,通过引物筛选和反应体系优化,建立阪崎肠杆菌快速检测的实时荧光环介导等温扩增(Loop-mediated isothermal amplification,LAMP)方法。以人工污染方式检验该方法在乳粉中阪崎肠杆菌检测灵敏度和稳定检测限,并对比实时荧光LAMP法和国标法在48份市售婴幼儿配方乳粉中的实际检测效果。结果显示,筛选ompA-2引物扩增效率最优,且与常见食源性致病菌无交叉反应;建立的实时荧光LAMP法对人工污染乳粉样品中的阪崎肠杆菌检测灵敏度、稳定检测限分别为10^2 cfu/g和10^3 cfu/g;48份市售乳粉样品进行增菌12 h,实时荧光LAMP法检测结果与传统国标培养结果一致。建立的实时荧光LAMP方法将为阪崎肠杆菌快速检测提供有力手段。
In order to achieve rapid detection of Enterobacter sakazakii in infant formula,an isothermal fluorescence amplification method was established to specifically detect Enterobacter sakazakii.Three sets of primers were designed for the specific conserved ompA gene of Enterobacter sakazakii.Through primer screening and reaction system optimization,a real-time fluorescent Loop-mediated isothermal amplification method for rapid detection of Enterobacter sakazakii was established.The artificial detection method was used to test the sensitivity and stable detection limit of Enterobacter sakazakii in milk powder,and the actual detection effect of real-time fluorescent LAMP method and national standard method in 48 commercial infant formula milk powder was compared.The results showed that the screening ompA-2 primer had the best amplification efficiency and no cross-reaction with common food-borne pathogens.The established real-time fluorescent LAMP method was used to detect the sensitivity and stable detection limit of Enterobacter sakazakii in artificially contaminated milk powder samples.It was 10^2 cfu/g and 10^3 cfu/g,respectively.48 commercial milk powder samples were enriched for 12 hours,and the results of real-time fluorescent LAMP method were consistent with the results of traditional national standard culture.The established real-time fluorescent LAMP method will provide a powerful means for rapid detection of Enterobacter sakazakii.