基于目标诱导链释放的高灵敏度信号放大技术的构建及食品中毒素检测研究

项目名称： 基于目标诱导链释放的高灵敏度信号放大技术的构建及食品中毒素检测研究

项目编号： No.21275085

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 数理科学和化学

项目作者： 混旭

作者单位： 青岛科技大学

项目金额： 80万元

中文摘要： 本课题拟开展基于目标诱导链释放的高灵敏度信号放大技术的构建及食品中毒素检测研究。课题拟利用目标诱导链释放和切口酶酶切作用，建立一个目标物产生多个信号单元的信号放大新技术，解决现有分析测定技术中信号放大倍数受"一对一化学计量识别"和酶切循环放大中目标物的适体需要特异性位点限制的科学问题，丰富和发展信号放大的理论基础；并以功能化的磁性微珠为捕获和分离载体，以负载ABEI（N-(4-氨基丁基)-N-乙基异鲁米诺）、识别作用和占位作用的DNA功能化化学发光纳米粒子为探针，以食品中毒素- - 赭曲霉毒素A为模式分析测定对象构建高灵敏度和高选择性食品中毒素的分析测定新方法。对实际样品中赭曲霉毒素A进行测定，利用色谱标准方法对照等对新方法进行评价。对建立的分析测定新方法进行拓展，将其发展成为一种共性检测技术，初步应用于其他食源性致病菌的检测。为高灵敏度食品安全检测提供有效的设计思路及实验依据。

中文关键词： 目标诱导链释放；信号放大；毒素；赭曲霉毒素A；化学发光

英文摘要： A novel signal amplified strategy based on target-induced strand release will be developed for the ultrasensitive detection of food toxins. The new signal amplified strategy, which one target produces multiple signal units, based on target-induced strand release coupling cleavage of nicking endonuclease will be firstly fabricated. This signal amplified strategy can solve the scientific problems of coefficient of signal amplification being restricted by one-to-one recognition reaction between target and signal unit and the needing of specific recognition of target aptamer in nicking endonuclease signal amplification. It also can enrich and develop the theory of signal amplification. The chemiluminescent probe will be prepared with gold nanoparticles, N-(4-aminobutyl)-N-ethylisoluminol (ABEI), recognition and occupation DNA. The Ochratoxin A (OTA), choosed as the interested target, will be sensitively and selectively detected with the developed signal amplified strategy combined with the magnetic bead capturing and separation. The feasibility and reliability of the developed method will be evaluated by the determination of OTA concentrations in agricultural commodities and HPLC controlled experiment. Generalization of this detection method for the others foodborne pathogens will also be studied. This study will pr

英文关键词： Target-induced strand release；Signal amplification；Food toxins；Ochratoxin A；Chemiluminescence