Research Center for Analytical Instrumentation, Institute of Cyber-Systems and Control, State Key Laboratory of Industrial Control Technology, Zhejiang University, Hangzhou 310027, PR China.
Research Center for Analytical Instrumentation, Institute of Cyber-Systems and Control, State Key Laboratory of Industrial Control Technology, Zhejiang University, Hangzhou 310027, PR China; College of Life Sciences, Zhejiang University, Hangzhou 310058, Zhejiang, PR China.
Biosens Bioelectron. 2018 Nov 30;120:144-152. doi: 10.1016/j.bios.2018.08.030. Epub 2018 Aug 17.
Digital polymerase chain reaction (dPCR) circumventing the external calibration and potentially providing absolute quantification of nucleic acids has become an increasingly popular manifestation of PCR in biological researches. However, currently reported or commercial dPCR devices are not suitable for applications in laboratories or zones with limited infrastructures, due to low function integration, cost-inefficiency, or weak mobility. Herein, in order to enable accurate DNA quantitative analysis in such situations, we have developed a smartphone-based mobile dPCR device integrated with thermal cycling control, on-chip dPCR, data acquisition, and result analysis. All the function units are automatically controlled using a customized Android software. The device is approximately 90 mm × 90 mm × 100 mm in size and about 500 g in weight, only costing about 320 dollars except the smartphone. Coupled with the self-priming dPCR chip previously developed by our lab, the device is able to accurately quantify down to 10 copies of the human 18 S ribosomal DNA fragment inserted in a plasmid. Comparing to the commercial QuantStudio™ 3D dPCR platform, our device achieves a comparable analytical accuracy. Besides, our device is capable of detecting single molecule of cancer biomarker gene CD147 in a low number of HepG2 cells. Therefore, our dPCR device as a low-cost, potable, and robust tool for highly accurate DNA quantitative analysis has a great potential in Point-of-care (POC) applications.
数字聚合酶链反应（dPCR）绕过了外部校准，并且有可能对核酸进行绝对定量，已成为生物学研究中聚合酶链反应越来越流行的表现形式。然而，由于功能集成度低、成本效益低或移动性弱，目前报告的或商业的 dPCR 设备并不适合在基础设施有限的实验室或区域中应用。在此，为了能够在这种情况下进行准确的 DNA 定量分析，我们开发了一种基于智能手机的移动 dPCR 设备，该设备集成了热循环控制、芯片上 dPCR、数据采集和结果分析。所有功能单元都使用定制的 Android 软件自动控制。该设备的尺寸约为 90mm×90mm×100mm，重量约为 500g，除了智能手机外，成本约为 320 美元。该设备与我们实验室之前开发的自启动 dPCR 芯片相结合，能够准确地对插入质粒的人类 18S 核糖体 DNA 片段的拷贝数进行定量，低至 10 个拷贝。与商业的 QuantStudio™3D dPCR 平台相比，我们的设备具有相当的分析准确性。此外，我们的设备能够在低数量的 HepG2 细胞中检测到单个癌症生物标志物基因 CD147 的存在。因此，我们的 dPCR 设备作为一种低成本、便携且强大的高精度 DNA 定量分析工具，在即时检测（POC）应用中具有巨大的潜力。
