Department of Electronic Engineering, Chang Gung University, Taoyuan 33002, Taiwan; Research Center for Informatics, Indonesian Institute of Sciences, Bandung 40135, Indonesia.
Department of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan; Division of Pediatric Infectious Disease, Department of Pediatrics, Chang Gung Memorial Hospital, Linkou 33305, Taiwan.
Biosens Bioelectron. 2017 Jun 15;92:186-191. doi: 10.1016/j.bios.2017.01.043. Epub 2017 Jan 21.
This study presents the first report on a label-free detection and rapid quantification method for human enterovirus 71 (EV71) using a portable surface plasmon resonance (SPR) system. The SPR sensor instrument was configured to run on low power in a miniaturized platform to improve the device portability for a wider application both in laboratories and in the field. A color tunable organic light emitting diode in red spectrum was attached on a trapezoidal prism for the disposable light source module. The SPR signal processing using integration area under the reflectivity curve is applied for optimum signal to noise ratio (SNR) enhancement. The major capsid protein VP1 of EV71 was selected as the biomarker target in the detection study. The experimental time required for the EV71 quantification was reduced from 6 days using the conventional viral plaque assay to several minutes using the proposed method. The study results establish a detection limit of approximately 67 virus particles per milliliter (vp/ml) of EV71 in a Dulbecco's modified Eagle's medium. The VP1 detection in the portable SPR biosensor had a detection limit of approximately 4.8pg/ml in the PBS buffer. Therefore, the proposed direct EV71 viral particle quantification method can be rapidly performed in real time, with high sensitivity and less labor and without assays or fluorescence.
本研究提出了一种使用便携式表面等离子体共振（SPR）系统对人肠道病毒 71（EV71）进行无标记检测和快速定量的方法，这在国际上尚属首次。该 SPR 传感器仪器配置为在小型化平台上以低功率运行，以提高设备的便携性，使其更广泛地应用于实验室和现场。一个红色光谱可调彩色有机发光二极管（OLED）被附加在一个梯形棱镜上，作为一次性光源模块。使用反射率曲线下的积分面积进行 SPR 信号处理，以增强最佳信噪比（SNR）。EV71 的主要衣壳蛋白 VP1 被选为检测研究中的生物标志物靶标。使用传统的病毒空斑测定法，定量 EV71 所需的实验时间从 6 天减少到使用所提出的方法的几分钟。该研究结果确定了在 Dulbecco 改良 Eagle 培养基中 EV71 的检测限约为每毫升 67 个病毒颗粒（vp/ml）。在 PBS 缓冲液中，便携式 SPR 生物传感器中 VP1 的检测限约为 4.8pg/ml。因此，所提出的直接 EV71 病毒粒子定量方法可以快速实时进行，具有高灵敏度、低劳动强度，且无需进行测定或荧光检测。
