点击反应构建可控型聚合物信号放大探针新策略及其在脓毒血症病原体快速筛查中的应用研究

脓毒血症是病原体感染引起的急性严重炎症反应，早期病原体诊断对有效治疗和提高病人存活率尤为关键。但现行的培养法筛查致病原耗时长且阳性率低，更高效的核酸检测法则由于依赖复杂的多重扩增技术而难以推广，所以亟需开发更简单的脓毒血症多重病原体检测新策略。申请人前期制备了金属配位聚合物及多官能团修饰的活化炔类聚合物光学材料，利用聚合后的放大效应实现其光-电转换效率或荧光信号的显著增强。同时，研究了活化炔的高效点击化学，这类反应条件温和、结构可控性高，在信号放大类聚合物的可控制备和修饰中有较大潜力。因此，本项目拟基于上述聚合物放大效应制备不需要扩增就能增强其电化学发光信号的核酸探针，并利用简单高效的点击反应提高探针的结构可控性，再结合多通道微流控芯片实现多种病原体的同步检测。通过该策略有望得到一种高效、高精度、条件要求简单的多重原体筛查新技术，为脓毒血症患者的早发现、早治疗提供数据及理论支持。

Sepsis is an acute severe inflammatory response induced by a broad spectrum of pathogens, with the mortality in severe cases higher than 25%. Rapid diagnosis of pathogens plays a key role in guiding effective treatment, and significantly improving the survival rate of patients. However, it takes several days for the clinical culture method to generate results, and the positive rate is low. Alternative technologies, for example, the specific nucleic acid detections of multi-pathogens are mainly built on multiplex PCR technology, which is too complex to be popularized. Thus, it is still urgent to develop a new strategy for the efficient identification of pathogens in sepsis. The applicant has successfully applicated the amplification effect of polymers in enhancing the photo-to-current conversion efficiency of metal complexes, and improving the emission of hyperbranched polymeric optical probes. At the same time, the highly efficient click chemistry of activated alkynes was also studied, which can be conducted under mild conditions with good structural controllability. This kind of reactions are potential to be used for the controllable preparation and modification of signal amplification polymers. Therefore, this project plans to constructing nucleic acid recognition probes with controllable structures and easy preparation procedures by simple and efficient click reactions. Their electrochemiluminescence signals can be enhanced by polymer amplification effect rather than PCR, which improves the speed and convenience of detection. In addition, these probes will be utilized on a multi-channel microfluidic chip to realize the simultaneous detection of multiple pathogens, in order to further simplify the steps and decrease the consumption of reagents. In summary, this strategy is expected to save the detection time and cost as much as possible, and provide a multiplex pathogen detecting technology with high efficiency, excellent accuracy, and simple steps, thereby providing data and theoretical support for early diagnosis and treatment of sepsis.