基于微流控纸基芯片的高灵敏度现场核酸检测方法研究及应用

项目名称： 基于微流控纸基芯片的高灵敏度现场核酸检测方法研究及应用

项目编号： No.21305162

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 数理科学和化学

项目作者： 慕轩

作者单位： 中国医学科学院基础医学研究所

项目金额： 25万元

中文摘要： 现场的核酸检测对于传染性疾病的防控和个体化医疗的发展具有十分重要的意义。但是核酸的传统检测方法对设备和操作要求较高，应用于现场有诸多困难。本项目拟针对检测灵敏度和前处理集成化等关键问题，充分利用微流控纸基芯片(μPAD)的特点和优势，系统研究和探讨高灵敏度现场核酸检测的新思路、新方法和新规律。本项目将重点提出适合纸基的高灵敏度核酸检测新思路，利用分支核酸(bDNA)技术的多重核酸杂交反应放大纸基上的核酸信号；提出利用疏水膜集成FTA elute卡的新方法，实现μPAD半自动全血前处理操作的集成化；结合动力学和扩散过程研究，揭示核酸杂交反应在纸基上的新规律。本项目还将研究缓冲液成份、反应温度、反应时间等对核酸杂交反应效率和速率的影响；检测全血中的疟疾诊断标志物(疟原虫18S rRNA)。本项目将为高灵敏度的现场核酸检测提供μPAD技术平台和理论指导，并促进μPAD在临床分子诊断中的广泛应用。

中文关键词： 微流控；临床诊断；丙型肝炎；囊性纤维化；纸基

英文摘要： Point-of-Care nucleic acids testing (POC NAT) is essential to the prevention and control of infectious disease as well as the development of personalized medicine. However, the traditional detection of nucleic acids normally requires strict operational environments, including complicated equipments and well-trained professionals, and thus is extremely difficult to apply to POC settings. This proposal would target the critical issues of sample preparation and signal detection of NAT, take full advantage of microfluidic paper-based analytical device (μPAD), and delve into new methodology, technology and mechanism with regard to high-sensitive POC NAT. We propose a new methodology to achieve high-sensitive NAT on paper matrix by utilizing multiple hybridizations of branched DNA (bDNA) to amplify the signal of nucleic acids; a new technology to integrate FTA card with μPAD by using a removable hydrophobic membrane, and perform semiautomatic whole blood preparations, such as cell lysis, RNA collection and release, on μPAD; and reveal new mechanisms of NAT on paper by combining kinetics of hybridization and diffusion of nucleic acids. We would also study the influence of buffer constituents, temperature and time on the effectiveness and rapidity of hybridization; employ μPAD to detect one diagnosis marker of malaria (

英文关键词： Microfluidics；Clinical Diagnosis；HCV；Cystic FIbrosis；Paper Matrix