基于多重PCR技术同时检测肉制品中6种动物源性成分  

Simultaneous determination of six animal-derived ingredients in meat product by multiple PCR method

作　　者：赵志勇[1] 朱少华 赵晓燕[1] 索玉娟[1] 陈爱亮[3] 李晓贝[1] 张艳梅 周昌艳[1] ZHAO Zhiyong;ZHU Shaohua;ZHAO Xiaoyan;SUO Yujuan;CHEN Ailiang;LI Xiaobei;ZHANG Yanmei;ZHOU Changyan(Institute for Agro-Food Standards and Testing Technology,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China;Engineering Construction Service Center,Ministry of Agriculture and Rural Affairs,Beijing 100125,China;Institute of Quality Standard and Testing Technology for Agro-products,Chinese Academy of Agricultural Sciences,Beijing 100081,China)

机构地区：[1]上海市农业科学院农产品质量标准与检测技术研究所,上海201403 [2]农业农村部工程建设服务中心,北京100125 [3]中国农业科学院农业质量标准与检测技术研究所,北京100081

出　　处：《上海农业学报》2021年第4期117-124,共8页Acta Agriculturae Shanghai

基　　金：上海市科委技术标准专项(16DZ0501800);上海市闵行区科委项目(2016MH210)。

摘　　要：为建立一种检测肉制品中6种动物源性成分(牛、羊、鸡、猪、鸭和驴等)的多重PCR方法,比较了3种DNA提取方法,设计筛选了6对引物并对其特异性进行验证,同时对影响多重PCR扩增效率的因素(引物比例、退火温度和镁离子浓度等)进行优化。结果表明:试剂盒法为最佳的DNA提取方法;多重PCR法最优反应条件:引物比例为牛∕羊∕鸡∕鸭∕猪∕驴=2∕1∕1∕0.5∕1∕1,退火温度为58.6℃,镁离子浓度为3 mmol∕L,各动物源性成分DNA最低检出限为0.1 ng。该方法具有特异性强、灵敏度高、简单快捷等优点,可用于肉制品中6种动物源性成分的同时检测。In order to develop a multiple PCR method for simultaneous determination of six animal-derived ingredients(cattle,sheep,chicken,pig,duck and donkey)in meat product.We compared three DNA extraction methods,designed and identified six high-specificity primers,optimized various factors affecting PCR amplification efficiency(primer ratio,annealing temperature and Mg^(2+)concentration).As a result,rapid kit method was selected as the best method of DNA extraction.The optimal conditions of multiple PCR method was as follows:primer ratio,cattle/sheep/chicken/pig/duck/donkey=2/1/1/0.5/1/1;annealing temperature,58.6℃;Mg^(2+)concentration,3 mmol/L.The limit of detection was 0.1 ng for six animal derived components.The major advantages of the developed method were good specificity,high sensitivity and simplicity,which could be proposed to apply in identification of animal derived ingredients in meat product.

关 键 词：肉制品 动物源性成分 六重PCR 检测 

分 类 号：TS2[轻工技术与工程—食品科学与工程] S81[农业科学—畜牧学]