利用LAMP技术快速检测羊肉制品中的鼠源性成分  

Rapid Detection of Murine-Derived Ingredients in Mutton Products Using Loop-Mediated Isothermal Amplification

作　　者：陈珍金 张璜 石磊[1] 叶蕾[1] 蔡凯贤 颜栋林 韦涛 谢耐珍 陈小聪 CHEN Zhenjin;ZHANG Huang;SHI Lei;YE Lei;CAI Kaixian;YAN Donglin;WEI Tao;XIE Naizhen;CHEN Xiaocong(Institute of Food Safety and Nutrition,Jinan University,Guangzhou 510632,China;Guangzhou Double Helix Gene Technology Co.Ltd.,Guangzhou 510000,China;Guangxi-ASEAN Food and Drug Safety Inspection and Testing Center,Nanning 530000,China)

机构地区：[1]暨南大学食品安全与营养研究院,广东广州510632 [2]广州双螺旋基因技术有限公司,广东广州510000 [3]广西东盟食品药品检验检测中心,广西南宁530000

出　　处：《食品科学》2021年第12期322-327,共6页Food Science

基　　金：河北省产业创新创业团队项目(205A7601D);广西重点研发计划项目(桂科AB18126084)。

摘　　要：为实现羊肉制品中鼠源性成分的定性检测,基于环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术,以鼠的线粒体基因(大鼠环氧化酶基因KP244683.1和小鼠的16S rRNA基因KY018919.1)为靶标,分别设计相应的LAMP特异性引物,通过优化反应条件,确定最佳LAMP反应体系。对混合模拟样品进行特异性、灵敏度和稳定性评估,应用LAMP和聚合酶链式反应检测方法分别对市售羊肉制品进行检测,对比分析检测结果。结果表明,本研究建立的LAMP方法特异性强,大鼠和小鼠的检出限分别为0.1%和0.5%,稳定性好,LAMP与聚合酶链式反应市售实际样本检测结果具有高度的一致性。因此,本实验建立的鼠源性成分LAMP检测方法操作简便、高效、准确,为鼠肉掺假快速诊断和基层诊断提供了新的选择,可作为羊肉制品中鼠源性成分的快速检测新方法。Loop-mediated isothermal amplification(LAMP)was employed for the qualitative detection of murine-derived ingredients in mutton products.Specific primers targeting the murine mitochondrial genes(rat COX gene KP244683.1 and mouse 16S rRNA gene KY018919.1)were designed,and the reaction conditions were optimized to determine the optimal reaction system.The specificity,sensitivity,and stability were evaluated using simulated mixed samples.LAMP was used to detect commercially available mutton products,and the obtained results were compared with those obtained with real-time PCR.The results showed that the method established in this study had good specificity and stability.The limits of detection(LODs)for rat-and mouse-derived ingredients were 0.1%and 0.5%,respectively.The results of LAMP were highly consistent with those of real-time fluorescent PCR.Therefore,the LAMP method proved simple,efficient,and accurate,which could provide a new choice for rapid diagnosis of murine meat adulteration and primary diagnosis,and could be used as a new rapid detection method for murine-derived ingredients in mutton products.

关 键 词：鼠源性成分 肉类掺假 环介导等温扩增技术 线粒体基因 羊肉制品 

分 类 号：TS207.3[轻工技术与工程—食品科学]