实时荧光pcr法鉴定食品中鲑亚科鱼成分.pdfVIP

194 2013, Vol.34, No.20 食品科学 ※分析检测 实时荧光PCR法鉴定食品中鲑亚科鱼成分 1,2 2, * 2 2 2 2 李进波 ，李 想 ，谌鸿超，潘良文，李 正 ，宋 青 (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) 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 m m 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%( / ) 。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。 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 Identification of Salmoninae in Foods by Real-Time PCR 1,2 2, * 2 2 2 2 LI Jin-bo ，LI Xiang ，CHEN Hong-chao ，PAN Liang-wen ，LI Zheng ，SONG Qing (1. School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China ； 2. Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200135, China) Abstract ：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 m m by mixing the fish of Salmo salar or 0.01% ( / ) Salvelinus alpinus with maize, chicken or crucian. Standard deviations and relative standard de