实时荧光PCR法鉴定食品中鲑亚科鱼成分  

Identification of Salmoninae in Foods by Real-Time PCR

作　　者：李进波[1,2] 李想[2] 谌鸿超[2] 潘良文[2] 李正[2] 宋青[2] 

机构地区：[1]华东理工大学生物工程学院,上海200237 [2]上海出入境检验检疫局,上海200135

出　　处：《食品科学》2013年第20期194-198,共5页Food Science

基　　金：上海市技术性贸易措施应对专项(12TBT011);"十二五"国家科技支撑计划项目(2011AA100807);上海出入境检验检疫局科技计划项目(HK001-2011)

摘　　要：根据鲑亚科鱼类生长激素基因保守序列设计特异性引物和探针,建立了一种鉴定食品中鲑亚科鱼类成分的实时荧光聚合酶链式反应(PCR)方法。结果表明:建立的检测方法高度特异于鲑亚科鱼类检测,鲑科的另两个亚科——白鲑亚科和茴鱼亚科样品均无扩增曲线。采用5种常见鲑亚科鱼品种可稳定检测到的最低DNA量为100pg;分别将大西洋鲑鱼和北极红点鲑鱼肉混入玉米、鸡肉和鲫鱼样品中测定的相对灵敏度为0.01%(m/m)。检测方法的重复性测试结果表明,Ct值标准偏差和相对标准偏差均在可接受范围内。运用建立的实时荧光PCR检测方法对25份市售实际样品进行测试,有4份标识含有"三文鱼"的样品未检测出鲑亚科鱼类成分。A real-time PCR method was developed to indentify the ingredients of Salmoninae in foods. Specific primers and probes were designed according to the conserved region of the growth hormone gene of Salmoninae. Results showed that the developed method was highly specific for Salmoninae detection. The detection limits were 100 pg using five species of Salmoninae as the templates. The relative detection limits were 0.01% （m/m） by mixing the fish of Salmo salar or Salvelinus alpinus with maize, chicken or crucian. Standard deviations and relative standard deviations of Ct values for five DNA concentrations were all in the acceptable range. Twenty-five commercial products labeled “containing salmon” were tested by the real-time PCR assay, and four samples were detected without Salmoninae ingredient. Thus, considering its high specificity, sensitivity and repeatability, we believed that the real-time PCR could be used to rapidly and accurately identify Salmoninae ingredient in foods.

关 键 词：实时荧光聚合酶链式反应 鲑亚科 生长激素基因 鉴定 

分 类 号：TS207.3[轻工技术与工程—食品科学]