基于液滴微流控技术的集成式数字PCR芯片  

Integrated Digital Polymerase Chain Reaction Chip Based on Droplet Microfluidic Technology

作　　者：刘聪 蒋克明[2] 周武平[2] 张涛[2] 张志强[2] 印晨宇[2] 黎海文 LIU Cong;JIANG Keming;ZHOU Wuping;ZHANG Tao;ZHANG Zhiqiang;YIN Chenyu;LI Haiwen(School of Engineering Science,University of Science and Technology of China,Hefei 230022,China;SuzhouInstitute of Biomedical Engineering and Technology,Chinese Academy of Sciences,Suzhou 215163,China)

机构地区：[1]中国科学技术大学工程科学学院,安徽合肥230022 [2]中国科学院苏州生物医学工程技术研究所,江苏苏州215163

出　　处：《新技术新工艺》2019年第5期1-6,共6页New Technology & New Process

基　　金：国家重大科学仪器设备开发专项(ZDYZ2013-1);苏州市科技计划项目(SYG201724);国家高技术研究发展计划项目(2015AA042402)

摘　　要：为了实现对核酸的高灵敏度检测,设计出一款新型的微滴数字聚合酶链式反应(ddPCR)芯片。该芯片由流动聚焦式微滴生成模块及后方的微滴储存腔构成。使用COMSOL软件对微滴生成及收集腔内液体流动进行仿真,验证芯片结构的合理性。使用软光刻技术及模压成型方法制得聚合物芯片并进行试验,结果表明,该芯片可以在70s内生成20000个直径为120μm(0.89nL)的微滴,微滴一致性较好(体积的变异系数为2.11%),且可以整齐、稳定地排列在收集腔内。应用该芯片直接进行原位PCR扩增及检测即可得出DNA检测结果。该芯片具有高灵敏度、高通量和高集成度的优势,避免了普通数字PCR的各种缺陷,如操作复杂、移液过程中的样品污染、样品损失、微滴破碎和融合。In order to achieve high sensitivity detection of nucleic acids, a chip of novel micro droplet digital polymerase chain reaction (ddPCR) was designed. The chip was composed of the flow-focusing and droplet-generating module, and the droplet storage chamber in the back. The COMSOL software was used to simulate the droplet formation and liquid flow in the collection chamber to verify the rationality of the chip structure. The polymer chip was obtained by soft lithography and compression molding, and the results showed that the chip can generate 20,000 droplets with a diameter of 120μm (0.89 nL) in 70 s. The droplet had good consistency (The coefficient of variation (CV) of volume was 2.11%), and can be arranged neatly and stably in the collection chamber. DNA detection results can be obtained by direct in situ PCR amplification and detection. The chip had the advantages of high sensitivity, high throughput, high integration, and avoided the defects of common digital PCR, such as complicated operation, sample contamination, sample loss, droplet breakage and fusion during pipetting.

关 键 词：微液滴 数字聚合酶链式反应 微流控芯片 核酸检测 COMSOL仿真 两相流水平集 

分 类 号：O655.9[理学—分析化学]