单核细胞增生李斯特氏菌LAMP试剂盒的研制及在食品检测中的应用  

Development and Application of Loop-mediated Isothermal Amplification Kit for the Detection of Listeria Monocytogenes in Foods

作　　者：张体银[1] 曹以诚 陈洵 郑晶[1] 黄晓蓉[1] 李志勇[3] 邵碧英[1] 黄嫦娇[1] 吴谦[1] 

机构地区：[1]福建出入境检验检疫局国家鳗鱼与食源性微生物检测重点实验室福建省检验检疫技术研究重点实验室,福州350001 [2]广州华峰生物科技有限公司,广州510663 [3]广东出入境检验检疫局,广州510623

出　　处：《中国食品学报》2013年第1期138-144,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家认监委行业标准修订计划项目(2008B160);福建省自然科学基金(2008J0247)

摘　　要：利用环介导等温扩增(1oop-mediated isothermal amplification,LAMP)技术建立食品中单增李斯特氏菌的检测方法。通过对反应条件的优化,组装单增李斯特氏菌LAMP检测试剂盒,并对其特异性、敏感性和适用性进行验证。试验结果显示:该试剂盒适用于食品中单增李斯特氏菌的快速检测,用环介导等温扩增法在65℃条件下,1h内可检出单增李斯特氏菌,其灵敏度达17CFU/mL,特异性为100%,无假阳性和假阴性出现。用该方法与国标方法分别检测120种样品中的单增李斯特氏菌,两种方法的符合率达100%。该试剂盒具有高效、特异性强和敏感性高等特点,可满足大批量样品快速筛选的要求。A Loop-mediated isothermal amplification （LAMP） method for detection of Listeria monocytogenes（LM） in food was established. After optimization of the reaction conditions of LAMP, including the concentrations of primers, lycine, dNTPs, and MgSO4, and the amplification temperature, LM-LAMP kit was assembled, meanwhile sensitivity, specificity and potential usefulness were evaluated in this paper. The results showed that the kit was available to detect the LM in foods. The LM could be rapidly and specifically detected by LAMP method within 1 hour at constant temperature of 65℃. The sensitivity of the kit to detect LM was at the level of 17 CFU/mL and the specificity of the approach was 100% without any false negative or positive action. 120 food samples were detected with the developed method and the National Standard method, and the results showed that the coincidence rate of these two methods was 100%. All of that demonstrated that the kit is effective, high sensitivity and specificity for the detection of LM, and available for screening of LM in a large number of food samples.

关 键 词：环介导等温扩增技术 单增李斯特氏菌 试剂盒 

分 类 号：TS207.4[轻工技术与工程—食品科学]