羊源性成分LAMP检测方法的建立  

Establishment of Loop-Mediated Isothermal Amplification for Detection of Mutton-derived Ingredients Based on Isothermal Amplification Platform

作　　者：徐淑菲[1] 孔繁德[1] 苗丽 林双庆[1] 林振基[1] XU Shufei KONG Fande MIAO Li LIN Shuangqing LIN Zhenji(Xiamen Entry-Exit Inspection and Quarantine Bureau, Xiamen 361026, China Henan Entry-Exit Inspection and Quarantine Bureau, Zhengzhou 450003, China)

机构地区：[1]厦门出入境检验检疫局,厦门361026 [2]河南出入境检验检疫局,郑州450003

出　　处：《经济动物学报》2016年第4期200-206,共7页Journal of Economic Animal

基　　金：厦门市科技计划项目(3502Z20154079);国家质检总局科技计划项目(2014IK089);福建省科技计划引导项目(2015Y0031)

摘　　要：根据羊cyt B基因设计3对特异性引物,运用GenieⅡ等温扩增荧光检测系统,以LAMP荧光检测方法为基础,建立了检测羊源性成分的LAMP方法,并对该方法进行了反应体系和条件的优化。试验结果表明:该方法操作简单、特异性好;灵敏度比普通PCR方法高100倍,比q PCR方法低10倍,达到5.928×10-3μg/μL;反应时间短,最快10 min即可完成反应。Three pairs of specific primers were designed according to the cytB gene of mutton genome. By using Genie Ⅱ real-time fluorescence isothermal amplification detection system and basing on the LAMP fluorescence detection, LAMP assay for detecting mutton-derived ingredients was established, the reaction system and condition were optimized. The results indicated that the method was easy to operate and very specific. It＇s sensitivity was 100-fold higher than that of conventional polymerase chain reaction and 10-fold lower than that of quantitative polymerase chain reaction and the detection limit was found to be 5.928×10-3μg/μL. The method was less time consuming, and the fastest total reaction time was approximately 10 minutes.

关 键 词：羊源性成分 特异性引物 等温扩增荧光检测系统 LAMP 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]