给-受体型共轭聚合物的设计、合成及其病原菌区分检测研究

Pathogen infection problem caused by bacteria, fungi and other pathogenic bacteria has become a global public health security issue. Thus, development of a simple, rapid and visual detection method of pathogenic bacteria is important for the diagnosis and prevention of pathogen infection. The object of this proposal is to develop a fluorescence resonance energy transfer (FRET) based optical approach for distinguishable detection of pathogenic bacteria which employ donor-acceptor conjugated polymers. Design and synthesize a series of α-mannose modified conjugated polymers with both donor and acceptor units, screening out polymers with better photophysical properties, concanavalin A lectin targeting ablility, and a wide range of fluorescent color change after bounding with concanavalin A lectin. These polymers could be able to bind with pathogenic bacteria through specific recognize ability of α-mannose and α-mannose-specific lectins located on the surface of these pathogenic bacteria. Due to the type and number difference of α-mannose-specific lectins on the surface of different pathogenic bacteria, differential detection of pathogenic bacteria could be obtained by analyzing fluorescent color change of the polymers which exhibit different FRET efficiency when interacting with different kinds of pathogenic bacteria. This method is simple, rapid, visible to the naked eye, and does not need any complicated equipment, toxic dyes or expensive labeled primers, so it has potential application value for the rapid visual detection of pathogenic microorganisms.

由细菌和真菌等病原菌引起的感染问题已经成为全球性的公共卫生安全问题，发展简单、快速、可视化的病原菌检测方法对病原菌感染的诊断及防治具有重要意义。本项目旨在发展一种基于给-受体型共轭聚合物的荧光共振能量转移机理的病原菌区分检测新方法，设计、合成一系列新型的α-甘露糖修饰的给-受体型共轭聚合物，从中筛选出光物理性质较好，能够靶向刀豆蛋白A凝集素（特异性结合α-甘露糖），并且在刀豆蛋白A凝集素的作用下荧光颜色的变化范围较宽的聚合物。该类聚合物能够靶向结合病原菌表面的特异性识别α-甘露糖的凝集素，由于不同种类病原菌表面能够识别α-甘露糖的凝集素的种类及数量不同，通过分析不同种类病原菌介导下聚合物荧光共振能量转移效率不同所致的荧光颜色变化，即可进行病原菌的区分。该方法简单、快速、肉眼可视，并且不涉及任何复杂仪器、剧毒染料及昂贵标记引物的使用，在病原微生物快速可视化检测领域具有潜在的应用价值。

给-受体型共轭聚合物及聚噻吩衍生物因其量子产率高、光稳定性好、能带可调、侧链可修饰等优点而备受关注。本项目立足给-受体型共轭聚合物及聚噻吩衍生物的设计制备、结构表征、性能及应用研究。通过“Suzuki偶联聚合”和“FeCl3 氧化聚合”两种方法制备了一系列给-受体型共轭聚合物及水溶性聚噻吩衍生物。研究表明，不同的给、受体单元的引入实现了聚合物光学性质的调控，不同的功能基团修饰赋予了聚合物特定的功能。侧链连接α-甘露糖的给-受体型共轭聚合物在不同种类病原菌表面聚集状态的差异使得给体单元到受体单元的荧光共振能量转移效率不同，聚合物的荧光颜色发生变化，从而实现了病原菌的可视化区分检测。本项目的成功实施不仅有利地促进了给-受体型共轭聚合物及聚噻吩衍生物的发展，而且进一步扩展了其在病原菌可视化区分检测、生物成像领域中的应用，并为后续的临床诊断及应用奠定了坚实的基础。在本项目的支持下，共发表sci论文6篇（均已标注），申请发明专利1项、实用新型专利1项。此外，相关成果受到企业的关注，签订聚合物设计合成技术开发项目2项，共计到款14万元。