羊乳中牛乳成分的直接实时环介导等温扩增检测  

Direct Real-time Loop Mediated Isothermal Amplification Method for Determination of Cow Milk Components in Goat’s Milk

作　　者：澹台玮 徐秦峰 马西亚 张文娟 TAN Taiwei;XU Qinfeng;MA Xiya;ZHANG Wenjuan(School of Food and Biological Engineering,Shaanxi University of Science&Technology,National R&D Center For Goat Dairy Products Processing 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：《中国乳品工业》2020年第1期38-41,46,共5页China Dairy Industry

基　　金：陕西省科技厅项目(2018KJXX-043、2019NY-126);陕西科技大学科研启动基金项目(2016QNBJ-01)

摘　　要：根据线粒体保守序列选取牛、羊特异性引物,建立无需核酸提取的直接实时环介导等温扩增(LAMP)技术,用于羊乳中掺入牛乳成分的检测。结果表明,所建立的LAMP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0.07 pg/μL和0.7 pg/μL,在掺假样品(即二元混合乳)中直接实时LAMP技术可检测到掺入1%的牛乳成分。通过对10种市售羊乳制品检测显示,直接实时LAMP方法与经DNA提取的实时LAMP方法检测结果一致。因此建立的直接实时LAMP技术无需核酸提取,样品经简单处理即可作为模板直接进行LAMP扩增,简化了样品前处理过程,缩短了检测时间,适用于羊乳制品中牛乳成分的快速检测。According to the mitochondria conserved sequence, bovine and goat specific primers were selected to establish a direct real-time Loop mediated isothermal amplification(LAMP) technique without nucleic acid extraction, which was used to detect the incorporation of cow milk in goat milk. The results showed that the established LAMP method had good specificity, no cross-reactions and false positives for other species, and the detection sensitivity of cow and goat was up to 0.07 pg/μL and 0.7 pg/μL. The detection limit for binary mixed milk was 1%. The detection of ten commercially available goat dairy products showed that the direct real-time LAMP was consistent with the conventional real-time LAMP method using extracted DNA. Therefore, the established direct real-time LAMP technology does not require nucleic acid extraction, and the sample can be used as a template for direct LAMP amplification after simple treatment, which simplifies the sample pretreatment process, shortens the detection time, and is suitable for rapid detection of cow milk components in goat dairy products.

关 键 词：环介导等温扩增 直接实时检测 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]