结直肠癌多种microRNA的单分子检测研究

Colorectal cancer is one of the leading causes of cancer mortality in the world. Establishing a sufficient and sensitive method for detecting microRNAs in the bloodstream and tissue is essential to the diagnosis, prognosis and chemotherapy of colorectal cancer. This project aims to develop a biological nanopore sensor which can selectively detect colorectal cancer-associated microRNAs at the single molecular level without the need for labels and amplification of the microRNA. The sensor, which uses a decoded oligonucleotide probe to generated a target-specific signature signal, can quantify and distingusih multiplex microRNAs. The analysis method and its related software will further developed for rapid recognization of signature signals from massive nanopore data. This ultra-sensitive approach will be ultimately applicated in simultaneous quantifying multiplex microRNAs in plasma and tissue samples from colorectal cancer patients.

结直肠癌是临床上发病率和死亡率最高的肿瘤之一。人体组织及血液中结直肠癌microRNA超灵敏检测方法的建立对于结直肠癌的早期诊断、分期、预后判断以及结直肠癌发病机制的研究等方面都具有重要意义。本项目针对实际样品中结直肠癌microRNA分子非标记、无需扩增、超灵敏、快速、可靠、低成本检测分析的需求，拟利用纳米孔单分子检测技术，设计制备一系列可与目标microRNA分子特异性结合的DNA探针分子，以期在单分子水平上获得每一种结直肠癌相关microRNA分子的纳米孔特征阻断信号；进一步开发相关microRNA纳米孔特征阻断信号快速识别分析方法及相应软件系统，从而最终构建应用于结直肠癌患者血清及组织中多种microRNA分子同时检测的纳米孔超灵敏检测分析系统。

本项目针对实际样品中结直肠癌microRNA分子非标记、无需扩增、超灵敏、快速、可靠、低成本检测分析的需求，设计功能化纳米孔道，获得单个疾病标志物分子特征阻断电流信号；构建了集多孔滤膜预处理系统及纳米孔道检测微池于一体的集成化纳米孔检测分析装置，实现了利用纳米孔道快速、直接检测血清样品；发展基于隐马尔科夫模型的纳米孔道大数据识别方法，在干扰信号中快速识别、还原、聚类纳米孔道单分子特征电信号，从而将所研制的纳米孔道分析装置系统应用于血清样品中结直肠癌MicroRNA分子的快速检测，并拓展应用于单个甲胎蛋白、单个淀粉样蛋白等的单分子电化学分析测量研究中。