免疫磁珠-环介导等温扩增快速检测巴氏杀菌乳中蜡样芽孢杆菌的四环素耐药基因tetL  

Rapid Detection of Bacillus Cereus with Tetracycline Resistance Gene tetL in Pasteurized Milk by Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification

作　　者：李雪利 汪思源 翟征远[1] 林建涵[2] 郝彦玲[1] LI Xueli;WANG Siyuan;ZHAI Zhengyuan;LIN Jianhan;HAO Yanling(College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083,China;College of Information and Electrical Engineering,China Agricultural University,Beijing 100083,China)

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083 [2]中国农业大学信息与电气工程学院,北京100083

出　　处：《中国乳品工业》2022年第4期52-56,共5页China Dairy Industry

基　　金：国家重点研发计划(2018YFC1604303)。

摘　　要：建立了免疫磁珠分离(IMS)结合环介导等温扩增(LAMP)检测巴氏杀菌乳中蜡样芽孢杆菌的四环素耐药基因tetL的方法。结果表明:经优化后,每微克链霉亲和素纳米磁珠与375 ng生物素标记的芽孢杆菌抗体偶联,在含有1×103 CFU/mL耐四环素蜡样芽孢杆菌的巴氏杀菌乳中,25μL免疫磁珠孵育1 h后,对蜡样芽孢杆菌BA117的捕获率为68.1%。该IMS-LAMP方法特异性高,2 h内对巴氏杀菌乳中耐四环素蜡样芽孢杆菌的检测灵敏度为24 CFU/mL。IMS-LAMP方法具有用时短,灵敏度高,操作简单等优点,可以有效检测巴氏杀菌乳中蜡样芽孢杆菌的四环素耐药基因。A method combining immunomagnetic separation(IMS)and loop-mediated isothermal amplification(LAMP)was developed to detect tetracycline resistance gene(tetL)of Bacillus cereus in pasteurized milk in this study.After optimization,25μL of streptavidin magnetic beads conjugated with 375 ng of biotin-labeled anti-Bacillus antibody presented a capture efficiency of 68.1%for 1×103 CFU/mL of the tetracycline resistant strain Bacillus cereus BA117 in pasteurized milk within 1 h.The IMS-LAMP method had a high specificity,and the detection limit was 24CFU/mL for tetracycline resistant Bacillus cereus in pasteurized milk within 2 hours.The IMS-LAMP assay provides a rapid,simple,and highly sensitive method for the detection of Bacillus cereus with tetracycline resistance gene tetL in pasteurized milk.

关 键 词：免疫磁珠 环介导等温扩增 蜡样芽孢杆菌 四环素耐药基因 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]