鱼肉制品中龙利鱼和巴沙鱼的鱼源性成分的双重实时荧光PCR快速检测法  

Dual real-time PCR method for rapid detection of sole fish and Pangasius bocourti-derived components in fish products

作　　者：李杰 钱云开 王建昌 LI Jie;QIAN Yunkai;WANG Jianchang(Linyi Inspection and Testing Center,Shandong Linyi 276000,China;Qinhuangdao Customs Technology Center,Hebei Qinhuangdao 066004,China;Shijiazhuang Customs Technology Center,Hebei Shijiazhuang 050000,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]石家庄海关技术中心,河北石家庄050000

出　　处：《中国食品卫生杂志》2022年第3期539-545,共7页Chinese Journal of Food Hygiene

基　　金：海关总署科研项目(2020HK229);河北省重点研发计划(21375501D)。

摘　　要：目的建立龙利鱼和巴沙鱼源性成分的双重实时荧光聚合酶链式反应(PCR)快速检测方法。方法根据13种龙利鱼的线粒体16S rRNA序列,设计通用引物和探针,根据巴沙鱼的线粒体cytb基因序列设计引物和探针,通过优化扩增反应体系进行双重实时荧光PCR扩增,达到快速检测产物的目的。结果本方法特异性良好,龙利鱼基因组DNA灵敏度可达到10^(-3)ng,在与鲤鱼粉混合的鱼肉制品中可检测,质量分数灵敏度达到1%。巴沙鱼基因组DNA灵敏度可达到10^(-4)ng,在与龙利鱼粉、大西洋鳕鱼粉混合的鱼肉制品中均可检测,质量分数灵敏度达到0.1%,在与米粉混合的鱼肉制品中可检测,质量分数灵敏度达到0.001%。结论本方法检测特异性高、所需时间短、灵敏度高,可满足鱼肉制品中龙利鱼真伪鉴别和巴沙鱼掺假的检测要求。ObjectiveTo establish a dual real-time PCR rapid detection method for sole fish and Pangasius bocourtiderived components.Methods16S rRNA gene sequence of 13 species of sole fish.Taq Man primers and probe sets were designed according to the mitochondrial cytb gene sequence of Pangasius bocourti,and the amplification reaction system was optimized for real-time PCR to achieve the purpose of rapid detection of products.Results could reach 10^(-3)ng sole fish DNA.Sole fish 16S rRNA gene could be detected in fish products mixed with carp meal,and the mass fraction sensitivity could reach 1%.The sensitivity could reach 10^(-4)ng Pangasius bocourti DNA.Pangasius bocourti cytb gene could be detected in fish products mixed with sole fish,atlantic cod meal and rice noodles,the mass fraction sensitivity of sole fish and atlantic cod meal could reach 0.1%,and the mass fraction sensitivity of rice noodles could reach 0.001%.Conclusion detection requirements of sole fish authenticity and Pangasius bocourti adulteration in fish meat products.

关 键 词：龙利鱼 巴沙鱼 舌鳎 实时荧光PCR 鱼肉制品 掺假 检测 

分 类 号：R155[医药卫生—营养与食品卫生学]