食品中驴源性成分环介导等温扩增检测方法的建立  

Establishment of Loop-Mediated Isothermal Amplification Method for Detection of Donkey Components in Foods

作　　者：柳毅[1] 尹斯雅[1] LIU Yi;YIN Siya(Baoding Product Quality Supervision and Inspection Institute,Baoding 071051)

机构地区：[1]保定市产品质量监督检验所,保定071051

出　　处：《食品工业》2020年第2期288-292,共5页The Food Industry

基　　金：河北省自然科学基金项目(C2008000216).

摘　　要：建立食品中驴源性成分环介导等温扩增检测方法,对市售食品进行驴源性成分检测分析。针对驴的线粒体ATPase 6基因设计特异性引物,优化LAMP反应条件,验证引物的特异性和检出限。在DNA模板1.0μL,内引物FIP/BIP各1.6μL,外引物F3/B3各0.2μL, dNTPs 2.5μL, Mg2+4μL, Bst DNA聚合酶缓冲液添加量2.0μL, Bst DNA聚合酶2.0μL体系下, LAMP法对食品中驴源性成分检测荧光可视法检测限为0.01%。LAMP方法检测食品中驴源性成分是切实可行的。与其他扩增技术相比,该方法具有操作简单、成本低、灵敏、快速的优势,适合于在基层单位进行推广应用。A method of loop-mediated isothermal amplification for detection of donkey derived components in foods was developed. Specific primers were designed for the mitochondrial ATPase6 gene of the donkey, and the LAMP reaction condition was optimized to verify the specificity and detection limit of the primers. Under the DNA template 1.0 μL, the primers FIP/BIP each 1.6 μL, the outer primers F3/B3 each 0.2 μL, dNTPs 2.5 μL, Mg2+4 μL and Bst DNA polymerase buffer solution 2.0 μL, the detection limit of fluorescence visible method for detection of donkey source in food was 0.01%. It was feasible to detect donkey components in foods by LAMP. Compared with other amplification technologies, it had the advantages of simple operation, low cost, sensitive and rapid. It was especially suitable for the popularization and application of the basic units in the developing countries.

关 键 词：环介导等温扩增技术 检测 食品 驴源性成分 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]