基于核酸适配体-纳米胶束的荧光多标记生物传感器检测8-羟基-2'-脱氧鸟苷

Development of high-sensitivity, high-throughput method for detecting urinary 8-OHdG is important for tobacco control and cigarette smoke exposure assessment. Combing the DNA/Dye multiple fluorescence labeling and micelle technique, we are aiming to develop a new aptamer-micelle based biosensor for small molecule detection. Firstly, utilize aptamer-PEG as hydrophilic group, long chain lipid as hydrophobic group to synthesis aptamer-micelle. Secondly, the multiple fluorescence labeling system will be proposed using the aptamer-micelle, which are playing two roles of binding dye with aptamer and doping dye with micelle. Thirdly, using the aptamer-micelle as identification and signal output, introducing the 8-OHdG-thrombin conjugate as bridge molecules, the biosensors will be developed for detection of urinary 8-OHdG with high-sensitivity. This project will focus on the proposal of the multiple-fluorescence labeling system with dual-function of the identification and detection, introducing the aptamer-micelle to small molecules analysis, improving the sensitivity and throughput of 8-OHdG biosensors. The innovation of this study is the development of detection platform for small molecules such as urinary 8-OHdG using aptamer-micelle as identification unit and signal amplification. This project may provide a simple, fast and low-cost method for urinary 8-OHdG analysis.

发展高灵敏、高通量的8-OHdG分析方法对评估人体烟气暴露和控制烟草流行具有重要意义。本项目拟结合荧光多标记以及纳米胶束技术，发展一种新型适配体-纳米胶束传感器用于小分子化合物的检测。首先，我们以凝血酶适配体-聚乙二醇共价结合物为亲水端，长链脂肪酸为疏水端制备适配体-纳米胶束。之后，利用适配体-纳米胶束可以结合（适配体）和包裹（纳米胶束）染料分子的特性，构建新型荧光多标记体系。最后，引入桥梁分子8-OHdG-凝血酶共价结合物，以适配体-纳米胶束为信号源和识别单元构建传感器实现尿液中8-OHdG的高灵敏检测。项目将重点解决兼具识别和检测双功能的荧光多标记体系构建，适配体-纳米胶束用于小分子化合物分析，提高检测灵敏度等关键问题。项目的开展有望为尿液中8-OHdG的检测提供一种简单、快速，低成本的新方法。

构建检测识别一体化的新型荧光多标记体系，实现荧光信号放大，高灵敏检测靶标物是本课题的主要目的，同时考虑到商业化应用前景，项目组开展了以下几项工作：（1）“抗体-核酸”识别检测一体化体系的构建，并用于烟气中苯并a芘的检测；（2）纳米胶束的合成、表征以及活性位点的引入；（3）基于纳米胶束的荧光多标记定量试纸条用于8OHdG检测；（4）新型荧光多标记体系用于微生物和烟草青枯病菌的检测。项目研制的8OHdG的荧光定量试纸条有望用于人体尿液的体外诊断，而开发的烟草青枯病试纸条已经在江西和福建进行了小规模应用。