核酸提取和多重PCR集成微流控芯片系统的研究

项目名称： 核酸提取和多重PCR集成微流控芯片系统的研究

项目编号： No.31500691

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 徐友春

作者单位： 清华大学

项目金额： 20万元

中文摘要： 核酸提取和多重PCR是分子生物学研究和分子诊断中的重要技术。但在应用中却常遇到以下问题：1）常规操作中，核酸提取和PCR是在开放环境中分开进行的，存在污染和RNA样品易降解的风险。2）多重PCR是一项多指标同步检测的技术，但多对引物之间的竞争和互相干扰使得体系构建非常困难。开发微流控芯片系统是解决以上问题的一个思路，目前已有在芯片上提取核酸并在单腔体PCR的报道，但尚无真正集成核酸提取和多重PCR的芯片系统被开发，因此这就成为本课题的目标。具体方案是在芯片的管道内壁修饰或填埋壳聚糖等核酸吸附物质，通过流体控制实现核酸的吸附-清洗-洗脱过程，而多重PCR的实现思路是利用微结构将PCR体系分割成多个独立的单重PCR体系，以多个单重PCR小体系来实现多重PCR。本课题拟构建的系统能从根本上降低核酸污染和降解的风险，解决多重PCR体系构建的难题，有望为相关研究和临床诊断开发更有效的核酸检测新技术。

中文关键词： 微流控芯片；生物分析；疾病诊断；检测

英文摘要： Nucleic acid extraction and multiplex PCR are the important technologies for molecular biology research and molecular diagnosis. However, there are some issues existed: 1) The nucleic acid extraction and multiplex PCR are conventionally performed in the open environment separately, which bring the risk of contamination and RNA degradation. 2) Multiplex PCR is an effective tool for simultaneous multiple target detection but is limited by the intrinsic interference and competition among primer pairs when it is performed in one reaction tube. The development of an integrated microfluidic analytical system is a solution to overcome the aforementioned problems. There are some reports about integration of nucleic acid extraction and PCR on chip, but multiplex PCR has not been integrated with nucleic acid extraction yet. Therefore, the aim of this project is to construct an integrated microfluidic system for on-chip nucleic acid extraction and multiplex PCR. The on-chip DNA extraction can be achieved based on the capture mechanism of nucleic acid adsorption to the surface of some materials, such as chitosan, and the extract-wash-release process can be accomplished with the external fluid control system. Besides, dividing a multiplex PCR into many single PCRs is a simple strategy to overcome the intrinsic interference of primer pairs. Due to the enormous advantages of microfluidics in flow manipulation and analysis, our microfluidic system has great potential to elevate the convenience of molecular diagnosis as well as reduce the contamination risk and cost. Through introducing microfluidics into nucleic acid extraction and multiplex PCR, we aim to provide a robust and convenient tool for molecular biology research and clinical applications.

英文关键词： microfluidic chip;bioanalysis;disease diagnosis;detection