一种用于数字PCR的微流控芯片的设计、制作和性能验证

Design,Fabrication and Performance Verification of a Microfluidic Chip for Digital PCR

为了获得可大批量制作并用于数字PCR技术的低成本的微流控芯片,采用注塑工艺制作了一种新型的微滴式数字PCR芯片。芯片材料为具有良好光学性能的环烯烃共聚物(COC),通道结构采用十字交叉型设计。将数字PCR试剂通过COC芯片可以产生直径为40μm的微滴(体积为16.75μL),通过在光学检测平台对比COC芯片和PDMS芯片生成的微滴的脉宽CV值和荧光CV值,证明了COC芯片具有良好的液滴生成效果;通过对人体EGFR基因质粒的绝对定量能力实验结果,表明芯片在101~106copies/μL范围内与DNA理论浓度具有很好的相关性。

In order to obtain a low cost microfluidic chip that can be made in large quantities and used in digital PCR tech-nology,a new microdrop digital PCR chip was fabricated by injection molding. The material of the chip is cycloolefin co-polymer （COC）with good optical properties. When the digital PCR reagent is passed through the COC chip,the droplets of 40 μm in diameter （volume 16.75PL）can be produced. By comparing the CV value of the pulse width and the fluorescence CV value of the droplets generated by the COC chip and the PDMS chip on the optical detection platform,it is proved that the COC chip has good droplet. The experimental results on the absolute quantitative ability of human EGFR gene plasmids showed that the chip had a good correlation with the theoretical concentration in the range of 10^1-10^6copies/μL.