面向循环外泌体miRNA分析的凝集增强荧光偏振信号放大新方法研究

The circulating exosome miRNAs are found to be significantly correlated with the occurrence and development of tumor and other diseases. They have the advantagesof good stability, high specificity and rich in body fluids, and can be used as one of the main liquid biopsy markers for early diagnosis of disease. This project focuses on the development of new method for separation and detection of exosome miRNAs with high sensitivity and specificity. We will develop new method for separation and identification of exosome miRNAs. We will also establish functional nanoprobes combine biological recognition ligation based on quantum dots, dye doped nanoparticles and fluorescent nanoparticles, and explore a variety of fluorescence polarization signal conversion strategies based on agglutination and signal amplification using nanomaterials. A series of convenient, sensitive and stable methods will be constructed based on enzyme free self-assembly and enzyme-catalyzed DNA signal amplification technology. This new method will be used for the determination of exosome miRNAs in human serum and urine samples. The project will provide a valuable platform for early clinical diagnosis, prognosis evaluation, and surgical effect monitoring. It is of great significance to improve the sensitivity and specificity of miRNA detection and promote the development of biopsy.

循环外泌体miRNA与肿瘤等疾病的发生、发展等过程密切相关，在体液中具有稳定性好、特异性强等特点，可作为重大疾病早期诊断的体液活检主要标记物之一。本项目针对新型肿瘤标志物循环外泌体及其miRNA的分离富集和高灵敏高特异性检测等问题，研究外泌体在不同体液中的提取、富集新技术；利用荧光量子点、染料掺杂纳米颗粒、荧光纳米簇等纳米材料，结合生物识别连接反应构建高效的新型功能纳米探针，探索基于凝集增强效应和纳米强化信号放大的荧光偏振信号转换新策略；通过无酶自组装和酶催化核酸信号放大机制，建立系列便捷、灵敏、稳定的外泌体miRNA检测新方法，实现其在血清、尿液等临床样品中的测定。该项目的实施，为实现癌症等疾病的早期诊断、预后及疗效预测、手术疗效监测及疾病复发预测等临床应用提供简便实用的技术平台。对于提高循环外泌体miRNA检测灵敏性和特异性，推动体液活检技术发展具有重要意义。

本项目基于核酸信号转换与信号放大技术、新型纳米材料的优良特性，发展了一系列具有原创性的荧光及荧光偏振和微流控芯片电泳分析新技术。研究开发了核酸信号放大结合纳米片增强的荧光与荧光偏振检测体系；核酸信号放大结合芯片电泳分离的激光诱导荧光与化学发光检测体系；基于碳基量子点的荧光生物成像分析新体系。建立了一系列生物传感新方法用于miRNA、蛋白质、生物活性分子、金属离子、农药等物质的高灵敏高选择性测定。在方法学研究的基础上，将提出的方法应用于人血清、肿瘤细胞、江河湖水等实际样品中相关物质的含量测定。这些新技术和新方法的提出，为生物医疗、环境监测等相关领域的研究提供了重要的技术支撑，对于提高痕量物质的精准检测水平和重大疾病的早期诊断具有重要的科学价值。本项目共发表论文14篇，在Anal. Chem.; Chem. Commun.; Analytica Chimica Acta; Talanta等 SCI影响因子5.0以上的期刊上发表论文8篇; 培养青年教师1人，博士研究生1人，硕士研究生7人。