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A rapid nucleic acid concentration measurement system with large field of view for a droplet digital PCR microfluidic chip
Lab on a Chip ( IF 6.1 ) Pub Date : 2021-07-28 , DOI: 10.1039/d1lc00532d
Jinrong Shen 1 , Jihong Zheng 1 , Zhenqing Li 1 , Yourong Liu 1 , Fengxiang Jing 2 , Xinjun Wan 1 , Yoshinori Yamaguchi 3 , Songlin Zhuang 1
Affiliation  

Droplet digital polymerase chain reaction (ddPCR) is an effective technique, with unparalleled sensitivity, for the absolute quantification of target nucleic acids. However, current commercial ddPCR devices for detecting the gene chip are time consuming due to complex image stitching. To address this issue, we propose a universal concentration determination system and realize one-time gene chip imaging with high resolution. All the functional units are controlled by self-developed software using the PyQt5 module in Python. Without stitching technology, images of the ddPCR chip (28 mm × 18 mm) containing 20 000 independent 0.81 nL micro chambers can be obtained in less than 15 seconds, which saves about 165 seconds. A white laser light source (2 mW cm−2) was employed as a substitute for the mercury lamp. Its wavelength matches well with typical fluorescent dyes (e.g., HEX, ROX and Cy5), and thus it can strengthen the fluorescence intensity for weak signals. The results also demonstrated that the correlation coefficient for the measured concentration and theoretical value was above 99%, by testing the ddPCR products with COVID-19 virus. Such a system can greatly reduce the time required for image acquisition and DNA concentration determination, and thus is able to speed up the lab-to-application process for ddPCR technology.

中文翻译:

一种用于液滴数字PCR微流控芯片的大视场核酸浓度快速测量系统

液滴数字聚合酶链反应 (ddPCR) 是一种有效的技术,具有无与伦比的灵敏度,可用于目标核酸的绝对定量。然而,由于复杂的图像拼接,目前用于检测基因芯片的商用ddPCR设备非常耗时。为了解决这个问题,我们提出了一种通用的浓度测定系统,并实现了高分辨率的一次性基因芯片成像。所有功能单元均由自主开发的软件控制,使用 Python 中的 PyQt5 模块。在没有拼接技术的情况下,可以在不到15秒的时间内获得包含20 000个独立的0.81 nL微室的ddPCR芯片(28 mm × 18 mm)的图像,节省约165秒。一个白色激光光源(2 mW cm -2) 被用作汞灯的替代品。它的波长与典型的荧光染料(HEX、ROX和Cy5)匹配良好,因此可以增强弱信号的荧光强度。结果还表明,通过用COVID-19病毒检测ddPCR产物,测得的浓度与理论值的相关系数在99%以上。这样的系统可以大大减少图像采集和 DNA 浓度测定所需的时间,从而能够加快 ddPCR 技术从实验室到应用的过程。
更新日期:2021-08-11
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