目标物转换策略构建电致化学发光高灵敏农残适体传感新方法研究

Electrogenerated chemiluminescence (ECL) technique has attracted considerable attentions in residual pesticide detection because of its remarkable intrinsic superiorities of high controllability and sensitivity. At present, the ECL technique for residual pesticide detection mainly depended on the reaction between pesticide molecule and active intermediate of co-reactant or the inhibiting principle of pesticide molecule on enzymes. But those methods showed some deficiency, such as low anti-interference ability and low sensitivity. This project intends to use aptamer as pesticide molecule identification element, with the help of DNA self-assembly technology and cyclic recognition amplification technology of DNA Walker, through target substitution strategy establishs novel ultra-sensitive ECL aptamer sensor for pesticide molecule detection (such as phorate, acetamiprid, chlorpyrifos and malathion) with high sensitivity and effectivity. More importantly, this proposed project not only provides a new idea based on ECL technology for the efficient and sensitive pesticide detection, but also revealed a new avenue and inspirations for trace analysis of other biotoxin small molecules, environmental pollutants, and food additives, which is of great significance for the development of ECL technology in the field of bioanalysis.

电致化学发光（ECL）分析技术由于其可控性和高的灵敏度，已成为农药残留检测的研究热点之一。目前ECL农药检测的方法主要基于农药分子与共反应试剂的活性中间体作用或农药分子对酶的抑制原理所构建，存在抗干扰能力差、灵敏度低等缺陷。本项目拟采用高特异核酸适体作为农药分子识别元件，通过目标物转换等策略，结合DNA自组装技术、DNA步行器循环识别放大技术，构建新型超灵敏ECL适体传感器，用于痕量残留农药分子（如甲拌磷、啶虫脒、毒死蜱、马拉硫磷等）的快速、高灵敏检测。本项目不仅为构建高效、灵敏的ECL农残检测方法提供新的思路，还为其他生物毒素小分子、环境污染物小分子、食品添加剂的痕量分析提供借鉴和启发，在食品安全、环境检测领域具有重要的理论意义和社会效益。

本项目合成了一系列高性能的电致化学发光（ECL）材料（如掺杂四-(4-氨基苯)乙烯的苝复合微晶（ETTA@Pe MCs）、苝四甲酸共价有机框架材料（PTCA-COF）、空心红荧烯银@金纳米立方(Rub-Ag@Au)、蒽-葫芦脲超分子纳米球 (Ant SNPs)等）。利用这些ECL材料，采用高特异核酸适体作为识别元件，通过目标物转换等策略，结合DNA自组装技术、DNA步行器循环识别放大技术，构建新型超灵敏ECL适体传感器。实现了农药分子啶虫脒、小分子DA、赭曲霉毒素A、miRNA-21等目标物的高灵敏检测，并将所制得的传感器初步应用到实际样品中相关目标物含量的灵敏监测。本项目不仅为构建高效、灵敏的ECL农残小分子检测提供了新的思路，还为构建新型的非核酸目标物的检测ECL体系提供了一种新途径。