一种基于多重实时荧光聚合酶链式反应熔解曲线分析的肉及肉制品掺假鉴别方法  

A Multiplex RT-PCR Melting Curve Analysis Method for Adulteration Identification in Meat and Meat Products

作　　者：周彤[1] 李家鹏[1] 李金春[1] 乔晓玲[1] 黄鑫[1] 陈曦[1] 杨君娜[1] 郭雅[1] LI Jiapeng;ZHOU Tong;LI Jinchun;QIAO Xiaoling;HUANG Xin;CHEN Xi;YANG Junna;GUO Ya(Beijing Key Laboratory of Meat Processing Technology, China Meat Food Research Center, Beijing 100068, China)

机构地区：[1]中国肉类食品综合研究中心肉类加工技术北京市重点实验室,北京100068

出　　处：《食品科学》2017年第12期217-222,共6页Food Science

基　　金：国家自然科学基金面上项目(31271877);"十二五"国家科技支撑计划项目(2014BAD04B05)

摘　　要：为实现肉及肉制品掺假快速鉴别,分别以猪、牛线粒体DNA的COXⅠ,绵羊、山羊、狗、狐狸、貉线粒体DNA的16S rRNA及鸡、鸭线粒体DNA的12S rRNA基因为靶位点,设计扩增产物熔解温度(T_m值)具有显著性差异的特异性引物,建立一种用于快速鉴别肉或肉制品中猪、牛、绵羊、山羊、鸡、鸭、狗、狐、貉9种源性成分的5重实时荧光聚合酶链式反应熔解曲线分析方法,通过特异性、灵敏度及市售样品的检测,对该方法进行检验和评价。结果表明:方法具有良好的特异性及灵敏度,单物种DNA检出限为0.001~1 ng,多物种混合DNA检出限均为0.1 ng,通过市售样品检测表明该方法可用于实际样品(包括生鲜样品和熟制样品)掺假的快速鉴别。This study was aimed to establish a rapid method for the identification of adulterated meat and meat products.Fivepairs of species-specific primers,yielding amplicons having significantly different melting temperatures(Tm),were designedtargeting the mitochondrial COXⅠgene of pig and bovine,the16S rRNA gene of sheep,goat,dog,fox and raccoon,and the12S rRNA gene of chicken and duck,respectively.The established multiplex RT-PCR-melting curve analysis method wasevaluated in term of specificity and sensitivity and was applied in the detection of commercial meat products.The methodshowed a good specificity and sensitivity,and was able to detect0.001–1ng of DNA from single species and0.1ng of DNAform mixed species.Thus,it can be used to test both raw and processed meat products.

关 键 词：肉类掺假 多重实时荧光聚合酶链式反应 熔解曲线分析 SYBR Green染料 动物源性成分 

分 类 号：TS207.7[轻工技术与工程—食品科学]