环介导等温扩增技术快速检测肉中单增李斯特菌  

Rapid Detection of Listeria monocytogenes in Meat by Loop-mediated Isothermal Amplification

作　　者：姜霞[1] 满朝新[2] 赵玥明 周文琦[1] 曲艳艳[1] 庞心怡[1] 姜毓君[1,2] 

机构地区：[1]东北农业大学食品学院/乳品科学教育部重点实验室,哈尔滨150030 [2]东北农业大学国家乳业工程技术研究中心,哈尔滨150086

出　　处：《中国食物与营养》2015年第11期52-56,共5页Food and Nutrition in China

基　　金：国家科技支撑计划课题(项目编号:2012BAD28B02;2013BAD18B11;2012BAD29B07)

摘　　要：通过建立的环介导恒温扩增（Loop-Mediated Isothermal Amplification,LAMP）方法以达到肉中单增李斯特菌快速、灵敏的检出。以特异性的hly A毒力基因作为靶基因,与6株非单增李斯特菌进行特异性试验,同时对不同培养浓度的单增李斯特菌进行了LAMP和PCR方法的灵敏度比较,进而应用LAMP法检测人工污染肉中的单增李斯特菌。结果表明：纯培养物中单增李斯特菌LAMP检出限为8.8×10-0CFU/m L,其灵敏度比普通PCR高100倍;在人工污染肉中单增李斯特菌的检出限为8.8×10-1CFU/m L,在1h内即可完成扩增反应。LAMP方法具备快速、特异、简单、灵敏度高等优势,在食品基质中单增李斯特菌的检测方面具有较好的应用前景。We established a rapid,sensitive method with high efficiency to detect the Listeria monocytogenes in meat by loop-mediated isothermal amplification（ LAMP）. The LAMP and PCR method were established to detect the reaction sensitivity and specificity of Listeria monocytogenes by the targeted virulence gene hly A. Listeria monocytogenes was detected in pure cultures and pork meat which was contaminated with Listeria monocytogenes with high specificity. Results showed that the LAMP assay could specifically detect 8. 8 × 100 CFU / m L Listeria monocytogenes in pure cultures,and its sensitivity was 100-fold than PCR. Besides,the reaction could be completed in l h. In the pork meat with the artificially contaminated cultures,the detection limit could reach 8. 8 × 101 CFU / m L. In conclusion,LAMP had great potential to be a promising detection method in food samples with rapid,sensitive,specifict and easy-to-operate advantages.

关 键 词：单增李斯特菌 环介导等温扩增 肉 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]