食品科学

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环介质等温扩增技术快速检测阪崎肠杆菌的方法

任立松,陈梦馨,陈 卓,马 龙   

  1. 1.新疆军区疾病预防控制中心,新疆 乌鲁木齐 830011;2.新疆医科大学公共卫生学院,新疆 乌鲁木齐 830011
  • 出版日期:2013-06-25 发布日期:2013-06-17

Rapid Detection of Cronobacter sakazakii Using a Loop-Mediated Isothermal Amplification (LAMP) Assay

REN Li-song,CHEN Meng-xin,CHEN Zhuo,MA Long   

  1. 1. Xinjiang Command Center for Disease Control and Prevention, Ürümqi 830011, China;
    2. College of Public Health, Xinjiang Medical University, Ürümqi 830011, China
  • Online:2013-06-25 Published:2013-06-17

摘要:

根据阪崎肠杆菌ompA基因序列,用生物软件设计环介质等温扩增技术(LAMP)特异性引物,优化反应条件,建立LAMP检测方法。分别用煮沸法、FTA卡、试剂盒法提取不同浓度细菌的DNA模板,用LAMP方法检出的最低浓度分别为104、103、103CFU/mL。用LMAP方法检测12株肠道菌,结果表明:其中7株阪崎肠杆菌分离株为阳性,其他为阴性。LAMP技术检测样品中阪崎肠杆菌与PCR方法相比,耗时短、结果易判定。

关键词: 阪崎肠杆菌, 环介质等温扩增技术, 检测

Abstract:

This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for rapid detection of
Cronobacter sakazakii. The gene sequences of ompA were aligned using the biologic software and four specific primers used for
LAMP were designed and the reaction condition was optimized. DNA templates from different cell concentrations were extracted by
boiling, FTA card and kit methods and detected by the LAMP method to obtain detection limits of 104, 103 CFU/mL and 103 CFU/mL,
respectively. Seven of 12 Cronobacter sakazakii isolates investigated were positive while all the others were negative. This method
may provide a more simple and rapid approach to detecting Cronobacter sakazakii in comparison with PCR method.

Key words: Cronobacter sakazakii, LAMP, detection