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  

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

作　　者：徐义刚 崔丽春[2] 李丹丹[3] 刘忠梅 李苏龙 张子群 张国财[2] 

机构地区：[1]黑龙江出入境检验检疫局检验检疫技术中心,黑龙江哈尔滨150001 [2]东北林业大学林学院,黑龙江哈尔滨150040 [3]海南出入境检验检疫局检验检疫技术中心,海南海口570125

出　　处：《食品科学》2012年第16期137-141,共5页Food Science

基　　金：国家质检总局科技计划项目(2008IK162);黑龙江省博士后基金项目(LBH-Z09)

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。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.

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 

分 类 号：R155.5[医药卫生—营养与食品卫生学]