The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan.
Sci Rep. 2018 Nov 30;8(1):17480. doi: 10.1038/s41598-018-35772-y.
MicroRNA expression analysis is an important screening tool for the early detection of cancer. In this study, we developed two portable three-dimensional microdevices for multiple singleplex RNA expression analysis by microRNA purification and qRT-PCR as a prototype for point-of-care testing. These microdevices are composed of several types of modules termed 'chemical IC chips'. We successfully reduced the heating area and fluorescence observation area, reduced the energy required for the reaction, and improved the portability of all systems in the devices. The purification microdevice could purify the microRNA from the sample using the FTA elute card system. The disposable reactor module mounted on both devices was easily fabricated by deforming a 100-μm-thick polypropylene film using an uncomplicated procedure. The qRT-PCR microdevice could perform reactions for samples of small volume. We purified microRNA from the HepG2 liver cancer cell line using the purification microdevice and confirmed the expression level of miR-224, which is a potential biomarker for liver cancer. Furthermore, we observed an increase in the fluorescence intensity when we performed qRT-PCR in the qRT-PCR microdevice. Therefore, the two developed microdevices show promise as a new portable tool for early cancer detection.
miRNA 表达分析是癌症早期检测的重要筛选工具。在这项研究中，我们开发了两种便携式三维微器件，用于通过 miRNA 纯化和 qRT-PCR 进行多重单重 RNA 表达分析，作为即时检测的原型。这些微器件由几种类型的模块组成，称为“化学 IC 芯片”。我们成功地减小了加热面积和荧光观察面积，降低了反应所需的能量，并提高了器件中所有系统的便携性。纯化微器件可以使用 FTA 洗脱卡系统从样品中纯化 microRNA。两个设备上都安装的一次性反应器模块可以通过使用简单的程序使 100-μm 厚的聚丙烯薄膜变形来轻松制造。qRT-PCR 微器件可以对小体积的样品进行反应。我们使用纯化微器件从 HepG2 肝癌细胞系中纯化 microRNA，并证实了 miR-224 的表达水平，miR-224 是肝癌的潜在生物标志物。此外，我们在 qRT-PCR 微器件中进行 qRT-PCR 时观察到荧光强度增加。因此，这两种开发的微器件有望成为用于早期癌症检测的新型便携式工具。
