基于实时荧光PCR的鱼肉制品中帝王鲑、银鲑、粉鲑源成分的快速检测及定量研究  

Real-time PCR method for rapid detection and quantitative study of king salmon,silver salmon,and pink salmon-derived components in fish products

作　　者：李杰 赵玉强 刘艳飞 沙玉彪 刘应炜 钱云开 LI Jie;ZHAO Yuqiang;LIU Yanfei;SHA Yubiao;LIU Yingwei;QIAN Yunkai(Linyi Inspection and Testing Center,Linyi 276000,China;Qinhuangdao Customs Technology Center,Qinhuangdao 066004,China;Qinhuangdao International Travel Health Center,Qinhuangdao 066004,China;Junan Inspection and Testing Center,Linyi 276000,China)

机构地区：[1]临沂市检验检测中心,山东临沂276000 [2]秦皇岛海关技术中心,河北秦皇岛066004 [3]秦皇岛国际旅行卫生保健中心,河北秦皇岛066004 [4]莒南县检验检测中心,山东临沂276000

出　　处：《食品与发酵工业》2025年第4期319-326,共8页Food and Fermentation Industries

基　　金：国家市场监督管理总局科技计划项目(2023MK079);河北省重点研发计划项目(213777109D);海关总署科研项目(2020HK229)。

摘　　要：建立一种利用实时荧光PCR技术快速检测帝王鲑、银鲑和粉鲑源性成分的方法。根据帝王鲑和银鲑的线粒体cytb基因序列以及粉鲑的COI基因序列设计Taq Man引物和探针组合,通过优化扩增反应体系进行实时荧光PCR扩增,达到快速检测帝王鲑、银鲑和粉鲑源性成分的目的。该方法特异性良好,耗时短,反应只需1 h;帝王鲑的基因组DNA灵敏度可达到10-1 ng;在与罗非鱼粉混合的鱼肉制品中可检测,质量分数灵敏度达到0.1%,质量分数误差≤±5.0%;银鲑的基因组DNA灵敏度可达到1 ng;在与罗非鱼粉混合的鱼肉制品中可检测,质量分数灵敏度达到1%,质量分数误差≤±7.7%;粉鲑的基因组DNA灵敏度可达到1 ng;在与罗非鱼粉混合的鱼肉制品中可检测,质量分数灵敏度达到1%,质量分数误差≤±2.8%。该方法检测特异性强、所需时间短、灵敏度高、质量分数误差小,可满足帝王鲑、银鲑和粉鲑真伪鉴别和定量分析的要求。This study aimed to establish a real-time PCR rapid detection method for king salmon,silver salmon,and pink salmon-derived components.TaqMan primers and probe sets were designed according to king salmon and silver salmon’s mitochondrial cytb gene sequence and pink salmon’s mitochondrial COI gene sequence,and the amplification reaction system was optimized for real-time PCR to achieve the purpose of rapid detection of king salmon,silver salmon,and pink salmon-derived components.This method had good specificity.This method had a short time and only took 1 hour to react.The sensitivity of king salmon could reach 10-1 ng DNA.King salmon cytb gene could be detected in fish products mixed with tilapia meal,and the mass fraction sensitivity could reach 0.1%,and the mass fraction error was less than or equal to±5.0%.The sensitivity of silver salmon could reach 1 ng DNA.Silver salmon cytb gene could be detected in fish products mixed with tilapia meal,and the mass fraction sensitivity could reach 1%,and the mass fraction error was less than or equal to±7.7%.The sensitivity of pink salmon could reach 1 ng DNA.Pink salmon COI gene could be detected in fish products mixed with tilapia meal,and the mass fraction sensitivity could reach 1%,and the mass fraction error was less than or equal to±2.8%.This method had high specificity,short time,high sensitivity and low mass fraction error,and could meet the authenticity identification requirements and quantitative analysis of king salmon,silver salmon,and pink salmon.

关 键 词：三文鱼 帝王鲑 银鲑 粉鲑 实时荧光PCR 检测 真伪鉴别 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程] O657.3[轻工技术与工程—食品科学与工程]