利用无标记分子信标及核酸染料TOTO-3检测转基因食品-食品科学.PDFVIP

174 2015, Vol.36, No.02 食品科学 ※安全检测 利用无标记分子信标及核酸染料TOTO-3 检测转基因食品 向东山 （生物资源保护与利用湖北省重点实验室，湖北民族学院化学与环境工程学院，湖北恩施 445000 ） 摘  要：利用无标记的分子信标，结合核酸染料TOTO-3 ，采用同步荧光分析法，建立一种高灵敏、高选择性 的花椰菜花叶病毒CaMV 35S启动子核酸片段（目标DNA ）的定量检测方法。在优化条件下，目标DNA浓度为 －10 －10 ∆I C 2 ×10 ～200 ×10 mol/L之间时，TOTO-3 的荧光强度（ ）与目标DNA浓度（ ）之间具有良好的线性关系， 其拟合的回归方程为∆I = 2.022 2 C＋18.411 1。方法检出限（3σ ）为1×10－10 mol/L 。该方法重复性好、灵敏度高、 选择性好、检出限低。利用该方法，结合不对称聚合酶链式反应技术，实现了对转基因食品的检测。 关键词：无标记分子信标；TOTO-3 ；荧光；转基因食品；检测 Detection of Genetically Modified Food with a Non-Labeled Molecular Beacon and a Nucleic Acid Dye TOTO-3 XIANG Dongshan (Key Laboratory of Biologic Resources Protection and Utilization of Hubei Province, School of Chemical and Environmental Engineering, Hubei Minzu University, Enshi 445000, China) Abstract: A highly sensitive and selective detection method for DNA of cauliflower mosaic virus (CaMV) 35S promoter (target DNA) was developed using a non-labeled molecular beacon (MB) and a nucleic acid dye TOTO-3. Under the optimum conditions, the fluorescence intensity of TOTO-3 exhibited good linear dependence on the quantity of the target -10 -10 I C DNA in the range of 2 ×10 –200 ×10 mol/L. The fitted regression equation was ∆ = 2.022 2 + 18.411 1, and the limit -10 of detection was 1 ×10 mol/L (3σ). The proposed method had a good precision, low detection limit, good selection and high sensitivity. The detection of genetically modified food could be achieved by this proposed method based on asymmetric polymerase chain react