羊乳中牛乳成分的可视化环介导等温扩增检测方法  

Establishment of visual loop-mediated isothermal amplification method for detection of bovine milk components in goat milk

作　　者：澹台玮 徐秦峰 张文娟 李艳妮 TANTAI Wei;XU Qin-feng;ZHANG Wen-juan;LI Yan-ni(School of Food and Biological Engineering,National R&D Center for Goat Dairy Products Processing Technology,Shaanxi University of Science&Technology,Xi′an 710021,China)

机构地区：[1]陕西科技大学食品与生物工程学院国家羊乳制品加工技术研发专业中心

出　　处：《陕西科技大学学报》2020年第1期71-75,80,共6页Journal of Shaanxi University of Science & Technology

基　　金：陕西省科技厅科技计划项目(2018KJXX-043,2019NY-126);陕西科技大学博士科研启动基金项目(2016QNBJ-01)

摘　　要：环介导等温扩增(LAMP)技术能够快速检测食品中的动物源性成分,但常用的有机检测染料稳定性差、Stoke′s位移小等导致荧光颜色区分不够明显.基于此,根据线粒体保守序列设计牛、羊特异性引物,同时采用核酸分子“光开光”钌(II)配合物[Ru(bpy)2(dppz)]2+作为荧光染料,建立了一种简便、快速、直观地检测羊奶制品中掺入牛源性奶成分的可视化LAMP方法.结果显示,该方法特异性好,对其他物种没有交叉反应及假阳性现象的发生,对牛、羊的检测灵敏度可达0.007 pg/μL和0.7 pg/μL,并能检测出羊乳中掺假0.1%的牛乳.因此,该方法可直接通过荧光目视比色判定检测结果,具有快速、直观、低成本等优点,为羊奶真实性现场快速检测提供技术支持.Loop mediated isothermal amplification(LAMP)technology can rapidly detect animal-derived components in food,but the commonly used organic detection dyes have poor stability and small Stoke′s shift,which makes it difficult to discriminate the fluorescence color of sample from blank.In this study,bovine and goat specific primers were designed based on mitochondrial conserved sequences,and the ruthenium(II)complex[Ru(bpy)2(dppz)]2+with nucleic acid molecule“light-on”effect was exploited as a fluorescent dye to establish a simple,rapid and intuitive visual LAMP method for the detection of bovine milk components in goat milk products.The results showed that the method was specific,and there was no cross-reactivity and false positive phenomenon in other species.The detection sensitivity of bovine and goat could reach 0.007 pg/μL and 0.7 pg/μL,respectively.The adulteration in goat milk could be detected 0.1%milk.Therefore,the method can directly determine the result by fluorescent visual colorimetry,which has the advantages of fast,intuitive,low cost,and may provide technical support for rapid on-site detection of goat milk authenticity.

关 键 词：环介导等温扩增 可视化检测 羊乳 牛乳 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]