针对精准分析前列腺癌分期指示因子的超灵敏双极电极-多色电致化学发光法研究

Prostate cancer stage indicators can reveal the pathogenesis and progression of prostate cancer, thereby, developing accurate, intuitive, low-cost and high-throughput indicator detection affords an important scientific significance towards the accurate and rapid judgment of stage and further improving the accuracy and efficiency of prostate cancer treatment. In view of the research hotspots and difficulties in accurate analysis indicators, this project intends to establish an ultra-sensitive bipolar electrode array-multicolor electrochemiluminescence detection chip to maximize the detection sensitivity of indicators by synthesizing three potentiometric-separated reagents corresponding to red, green and blue, will achieve multicolor covering the entire visible light area, and combining with multiple current amplification strategies. On this basis, the integration of automated injection, highly-selective separation and ultra-sensitive detection systems is used to construct an integrated microfluidic analysis system for detecting multiple indicators accurately and visually, and prepare the standard precision swatches of multiple indicators. Finally, this above system will be used for clinical analysis, and the obtained images will be further compared with standard swatches to judge the prostate cancers stage accurately, which could lay a foundation for precise medical treatment of prostate cancer. The implementation of this project not only provides methodological support and theoretical basis for prostate cancer stage detection, but also provides a solution to other cancer stage detection, which has a broad application foreground.

前列腺癌分期指示因子可揭示前列腺癌的病变发生与进程，发展精准、直观、低成本和高通量的指示因子检测方法对于准确快速判断分期，提高前列腺癌治疗的精准性和效率具有重要的科学意义。针对精准可视化同时分析多指示因子这一大研究热点和难点，本项目拟通过合成三种电位分开的红光、绿光和蓝光试剂，实现混合光颜色覆盖整个可见光区域，并耦合多重电流放大策略，建立超灵敏的多色电致化学发光传感-双极电极阵列芯片，最大限度提高指示因子的检测灵敏度。在此基础上，整合自动化进样、高选择性分离和超灵敏检测系统，构筑一体化微流控分析系统，精准和可视化联检多指示因子，并制备多指示因子的标准精密色板。最后将上述系统用于临床分析，将获得的图像对照标准色板，准确快速评判前列腺癌分期，为前列腺癌“精准医疗”奠定基础。本项目的实施不仅为前列腺癌分期检测提供方法学支持和理论依据，还为其他癌症分期检测提供一种解决方案，具有广阔的应用前景。

“闭合式”BPE结构可将不同储液池中的ECL反应分开，因此常被用来构建比率-可视化ECL生物传感器，然而，目前这种类型的生物传感器对miRNA还不能实现超灵敏检测，主要原因是具有高量子产率和可视化光强的ECL试剂选择有限，以及流经BPE的电流不足。因此本项目在合成高量子产率和可视光强的ECL试剂，及发展高效的扩增技术来极大地提高可视BPE-ECL生物传感器的灵敏度方面开展了一系列的工作，包括：1、利用高量子产率和低毒性的BNQDs涂覆在闭合式BPE阴极作为ECL发射体，阳极用Ir(df-ppy)2(pic)材料进行校准，再通过BNQDs和Firpic的ECL比率来测量miRNA-21的浓度。2、基于可视化BNQDs构建了一个恒定电阻集成的闭合式双极电极的超灵敏“颜色开关“ECL生物传感器，实现超灵敏检测前列腺癌标记物miRNA-141。3、建立了一种基于WO3-x NDs/S-BNQDs的SPC-ECL和AuNPs/S-BNQDs的ECL-RET协同效应的新型传感策略，以实现对miRNA-141的高灵敏度检测。