肉串制品中猪牛羊源性成分多重荧光PCR检测方法研究  

Study on Detection Method for Pig,Cow and Sheep Derived Components by Multiple Fluorescence PCR in Meat Skewer Products

作　　者：姚艳玲 周宇东 王文宇 朱洪亮 陈婷 翁光灿 孙世元 YAO Yan-ling;ZHOU Yu-dong;WANG Wen-yu(Jiaxing Testing Institute for Food,Drug and Product Quality Control,Jiaxing,Zhejiang 314050)

机构地区：[1]嘉兴市食品药品与产品质量检验检测院,浙江嘉兴314050

出　　处：《安徽农业科学》2024年第18期184-187,共4页Journal of Anhui Agricultural Sciences

基　　金：浙江省市场监督管理局雏鹰计划培育项目(CY2023322)。

摘　　要：[目的]建立畜类肉串产品中常见的黄牛、绵羊和猪源性成分的多重荧光PCR检测方法,实现肉串制品中动物源性成分的快速鉴别。[方法]比较磁珠法和柱膜法对肉糜组织中DNA的提取效果,优化实时荧光PCR多重检测方法,比较58、60℃2个退火温度下黄牛、绵羊、猪源性成分扩增效果。[结果]试验采用的2种DNA提取方法效果相似,DNA溶液的A_(260)/A_(280)值均能满足PCR检测要求;58℃为最优退火温度,此时黄牛、绵羊、猪源性成分均能有效扩增,且最低检出的DNA浓度均为10^(-3) ng/μL。方法中用到的引物和探针特异性较好,均未产生非特异性扩增。[结论]试验建立的黄牛、绵羊和猪源性成分的多重PCR检测方法能为肉制品中动物源性成分的快速定性分析提供方法参考。[Objective]To establish a multiple fluorescence PCR detection method for common cattle,sheep and pig derived components in animal meat skewer products,and to achieve rapid identification of animal derived components in meat skewer products.[Method]Compared the extraction effect of DNA from meat mince tissue using magnetic bead method and column membrane method,optimized the real-time fluorescence PCR multiple detection method,and compared the amplification effect of cattle,sheep,and pig derived components at two annealing temperatures of 58 and 60℃.[Result]The two DNA extraction methods used in the experiment had similar effects,and the A 260/A 280 values of the DNA solution could meet the requirements of PCR detection.58℃was the optimal annealing temperature,at which point the cattle,sheep and pig derived DNA could be effectively amplified,and the lowest detected DNA concentration was 10^(-3) ng/μL.The primers and probes used in the method had good specificity and had not produced non-specific amplification.[Conclusion]The multiplex PCR detection method established by the application scope experiment for cattle,sheep and pig origin can provide a method reference for rapid qualitative analysis of animal origin components in meat products.

关 键 词：黄牛 绵羊 猪 源性成分 实时荧光多重PCR 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]