Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 575-0871, Japan; Eco Solutions Company, Panasonic Corporation, 1048 Kadoma, Kadoma, Osaka 571-8686, Japan.
Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 575-0871, Japan.
Biosens Bioelectron. 2015 Dec 15;74:725-30. doi: 10.1016/j.bios.2015.07.009. Epub 2015 Jul 9.
Polymerase chain reaction (PCR)-based genetic testing has become a routine part of clinical diagnoses and food testing. In these fields, rapid, easy-to-use, and cost-efficient PCR chips are expected to be appeared for providing such testing on-site. In this study, a new autonomous disposable plastic microfluidic PCR chip was created, and was utilized for quantitative detection of pathogenic microorganisms. To control the capillary flow of the following solution in the PCR microchannel, a driving microchannel was newly designed behind the PCR microchannel. This allowed the effective PCR by simply dropping the PCR solution onto the inlet without any external pumps. In order to achieve disposability, injection-molded cyclo-olefin polymer (COP) of a cost-competitive plastic was used for the PCR chip. We discovered that coating the microchannel walls with non-ionic surfactant produced a suitable hydrophilic surface for driving the capillary flow through the 1250-mm long microchannel. As a result, quantitative real-time PCR with the lowest initial concentration of human, Escherichia coli (E. coli), and pathogenic E. coli O157 genomic DNA of 4, 0.0019, 0.031 pg/μl, respectively, was successfully achieved in less than 18 min. Our results indicate that the platform presented in this study provided a rapid, easy-to-use, and low-cost real-time PCR system that could be potentially used for on-site gene testing.
聚合酶链式反应（PCR）为基础的基因检测已成为临床诊断和食品检测的常规部分。在这些领域中，快速、易于使用且具有成本效益的 PCR 芯片有望用于现场提供此类检测。在这项研究中，创建了一种新的自主式一次性塑料微流控 PCR 芯片，并用于定量检测病原体微生物。为了控制 PCR 微通道中后续溶液的毛细流动，在 PCR 微通道后面新设计了一个驱动微通道。这使得通过简单地将 PCR 溶液滴入入口即可有效地进行 PCR，而无需任何外部泵。为了实现一次性使用，成本效益高的注塑成型环烯烃聚合物（COP）被用于 PCR 芯片。我们发现，将非离子表面活性剂涂覆在微通道壁上可产生合适的亲水表面，以通过 1250mm 长的微通道驱动毛细流动。结果，成功地在不到 18 分钟的时间内实现了最低初始浓度为 4、0.0019、0.031pg/μl 的人、大肠杆菌（E.coli）和致病性大肠杆菌 O157 基因组 DNA 的定量实时 PCR。我们的结果表明，本研究中提出的平台提供了一种快速、易于使用且低成本的实时 PCR 系统，可用于现场基因检测。
