食品中鲑亚科鱼成分鉴定方法研究.pdfVIP

2013-03-13 食品中鲑亚科鱼成分鉴定方法研究 1,2 2* 2 2 2 2* 李进波 ，李想 ，谌鸿超 ，李正 ，宋青 ，潘良文 (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(growth hormone，GH)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 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。对建立的实时荧光 PCR 方法的检测下限（LOD ） 进行了测试。结果表明，采用 6 种常见鲑亚科鱼品种可稳定检测到的最低 DNA 量为 100 pg；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 LOD 为 0.01 % (W/W) 。检测方法的可重复性测 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。因此，本研究建立的实 时荧光 PCR 检测方法特异性好，灵敏度高，可重复性好，能快速、准确鉴定食品中鲑亚科鱼类成分。 关键词：实时荧光 PCR ；鲑亚科；生长激素基因；鉴定 Identification of Salmoninae in Food 1,2 2* 2 2 2 2* LI Jin-bo , LI Xiang , Chen Hong-chao , Li Zheng , Song Qing , PAN Liang-wen (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 ：To indentify the ingredients of Salmoninae in food, specific primers and probe for real-time PCR were designed according to the conservation region of growth hormone gene of Salmoninae. Results showed that the developed method was highly specific for Salmoninae detection. The detection limits of the esteblished method were 100 pg using six species of Salmoninae as the templates. The relative detection limits were 0.01% (w/w) by mixing the fish of Salmo Salar or Salvelinus alpinus with maize, chicken or crucian. Standard deviations and relative standard deviations of Ct values of five concentrations of DNA were all in the acceptable range. Twenty five commercial products which was labeled “containing s