State Key Laboratory of Applied Optics, Changchun Institute of Optics, Fine Mechanics and Physics (CIOMP), Chinese Academy of Sciences, Changchun 130033, China.
University of Chinese Academy of Sciences (UCAS), Beijing 100049, China.
Sensors (Basel). 2020 Apr 28;20(9):2492. doi: 10.3390/s20092492.
In recent years, the development of polymerase chain reaction (PCR) technology has focused on digital PCR, which depends on the microfluidics. Based on continuous-flow microfluidic technology, this paper designed a miniaturized digital PCR amplification system, and greatly reduced the area required for microdroplet generation and reaction. The core rod. made of polydimethylsiloxane (PDMS), was combined with the Teflon tube to form 3D microfluidics, which requires only one heating source to form the temperature difference required for gene amplification. Only two 34 g needles can form and transmit micro-droplets in a 4-fold tapered Teflon tube, which is the simplest method to generate digital PCR droplets as far as we know, which allows the microdroplet generation device to be free from dependence on expensive chips. A complementary metal oxide semiconductor (CMOS) camera was used as a detection tool to obtain fluorescence video for the entire loop area or a specified loop area. In addition, we developed a homebrew for automatic image acquisition and processing to realize the function of digital PCR. This technique realizes the analysis of clinical serum samples of hepatitis B virus (HBV) and obtained the same results as real-time quantitative PCR. This system has greatly reduced the size and cost of the entire system, while maintaining a stable response.
近年来，聚合酶链反应（PCR）技术的发展集中在数字 PCR 上，它依赖于微流控技术。本文基于连续流微流控技术，设计了一种小型化的数字 PCR 扩增系统，大大减少了微滴生成和反应所需的面积。核心棒由聚二甲基硅氧烷（PDMS）制成，与特氟隆管结合形成 3D 微流控，仅需一个加热源即可形成基因扩增所需的温差。仅用两根 34 g 针就可以在 4 倍锥形特氟隆管中形成和传输微滴，这是迄今为止我们所知的最简单的生成数字 PCR 微滴的方法，这使得微滴生成设备能够摆脱对昂贵芯片的依赖。互补金属氧化物半导体（CMOS）相机用作检测工具，以获取整个循环区域或指定循环区域的荧光视频。此外，我们开发了一个自制的自动图像采集和处理软件，以实现数字 PCR 的功能。该技术实现了对乙型肝炎病毒（HBV）临床血清样本的分析，得到了与实时定量 PCR 相同的结果。该系统大大缩小了整个系统的尺寸和成本，同时保持了稳定的响应。
