克罗诺杆菌属快速检测体系的建立  

Establishment of a Rapid Detection System for Cronobacter spp.

作　　者：王丹丹 李莉[1] 杨艳歌[1] 刘鸣畅[1] 王洪越 吴淑清 袁飞[1] 吴亚君[1] WANG Dandan;LI Li;YANG Yange;LIU Mingchang;WANG Hongyue;WU Shuqing;YUAN Fei;WU Yajun(Chinese Academy of Inspection and Quarantine,Beijing 100176,China;College of Food Science and Engineering,Changchun University,Changchun 130022,China)

机构地区：[1]中国检验检疫科学研究院,北京100176 [2]长春大学食品科学与工程学院,吉林长春130022

出　　处：《食品科学》2021年第16期286-292,共7页Food Science

基　　金：“十三五”国家重点研发计划重点专项(2018YFC1603606);中国检验检疫科学研究院基本科研业务费项目(2019JK003)。

摘　　要：以克罗诺杆菌属具有代表性的2株模式菌株(丙二酸盐阳性克罗诺杆菌和阪崎肠杆菌)为实验对象,建立了一套集一步法DNA提取和快速实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)法检测为一体的速测技术体系。通过优化,建立的快速real-time PCR程序可将扩增时间由普通程序的82 min缩短至27 min 18 s。建立的一步快速DNA提取流程,取样后仅需12 min热循环反应即可直接获取DNA,简便快捷。与传统方法的对比结果显示,快提法与传统煮沸法对纯菌液和人工污染培养前6 h的样品灵敏度一致;对人工污染样品培养7、8 h时,快检法的灵敏度比传统方法低1个数量级。本研究建立的微生物速测技术快速、简便且具有较高灵敏度,在口岸食源性病源微生物现场快速检测方面具有较好的应用前景。In this article,two typical model strains of Cronobacter spp.(Cronobacter malonaticus and Enterobacter sakazakii)were used to establish a rapid detection system integrating one-step DNA extraction and rapid real-time polymerase chain reaction(real-time PCR)for Cronobacter spp..The amplification time of the optimized fast real-time PCR was only 27 min 18 s compared to 82 min for the common procedure.In the one-step DNA extraction procedure,DNA could be directly obtained after thermal cyclic reaction for only 12 min.Compared with the traditional boiling method,the rapid detection method showed consistent sensitivity to pure culture and artificially contaminated samples cultured for 6 h.The sensitivity of the new method to artificially contaminated samples cultured for 7 and 8 h was one order of magnitude lower than that of the conditional method.The method presented in this study is fast,simple,and highly sensitive,and has a good application prospect in the rapid on-site detection of foodborne pathogens at ports.

关 键 词：微生物污染 食品安全 快速实时聚合酶链式反应 克罗诺杆菌属 快检技术 

分 类 号：TS207.4[轻工技术与工程—食品科学]