食品科学 ›› 2012, Vol. 33 ›› Issue (16): 137-141.

• 分析检测 • 上一篇    下一篇

食品中单核细胞增生李斯特菌DNA环介导恒温扩增快速检测方法的建立

徐义刚1,崔丽春2,李丹丹3,刘忠梅1,李苏龙1,张子群1,张国财2,*   

  1. 1.黑龙江出入境检验检疫局检验检疫技术中心 2.东北林业大学林学院 3.海南出入境检验检疫局检验检疫技术中心
  • 收稿日期:2011-07-06 修回日期:2012-07-04 出版日期:2012-08-25 发布日期:2012-09-07
  • 通讯作者: 徐义刚 E-mail:yigangxu_china@yahoo.com.cn
  • 基金资助:

    国家质检总局科技计划项目;黑龙江省博士后基金

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Listeria monocytogenes in Foods

  • Received:2011-07-06 Revised:2012-07-04 Online:2012-08-25 Published:2012-09-07

摘要: 基于单核细胞增生李斯特菌胞壁质水解酶iap基因,设计两对特异性引物,利用DNA环介导恒温扩增(loop-mediated isothermal amplification,LAMP)技术,以扩增副产物焦磷酸镁实时浊度为判定标准,建立食品中单核细胞增生李斯特菌LAMP快速检测方法。结果显示,本LAMP方法特异性强,经过对29株细菌进行检测,所试单核细胞增生李斯特菌均为LAMP阳性,其他菌株为阴性;本LAMP方法对单核细胞增生李斯特菌纯培养菌的检测灵敏度为8CFU/管,对污染食品中单核细胞增生李斯特菌的检测灵敏度为12CFU/管。本研究建立的LAMP检测方法简便快速、结果判断直观。

关键词: 单核细胞增生李斯特菌, iap基因, 环介导恒温扩增, 快速检测 

Abstract: According to the iap gene of Listeria monocytogenes, two pairs of specific primers were designed, and then a rapid loop-mediated isothermal amplification (LAMP) assay for detecting Listeria monocytogenes in foods was developed using real-time turbidity of the amplification byproduct magnesium pyrophosphate as the positive criterion. Twenty-nine bacterial strains were used to evaluate the specificity of the LAMP method. Our results showed that all tested Listeria monocytogenes were positive, while other strains were negative to LAMP detection, suggesting that this LAMP method was highly specific to the target bacteria. The sensitivity for cultivated Listeria monocytogenes and its contaminated foods were was 8 CFU and 12 CFU per test tube, respectively. The LAMP method developed in this study provides a sensitive, rapid and simple approach for the detection of Listeria monocytogenes.

Key words: Listeria monocytogenes, iap gene, LAMP, rapid detection

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