Institute of Applied Mechanics, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, 10617, Taiwan; Graduate Institute of Nanomedicine and Medical Engineering, International PhD Program in Biomedical Engineering, School of Biomedical Engineering, International PhD Program in Biomedical Engineering, International PhD Program for Cell Therapy and Regeneration Medicine Taipei Medical University, 250 Wuxing St, Taipei, 11031, Taiwan; Graduate School of Biomedical & Health Sciences, Hiroshima University, Kausmi 1-2-3, Minami-ku, Hiroshima, 734-8553, Japan.
Graduate Institute of Nanomedicine and Medical Engineering, International PhD Program in Biomedical Engineering, School of Biomedical Engineering, International PhD Program in Biomedical Engineering, International PhD Program for Cell Therapy and Regeneration Medicine Taipei Medical University, 250 Wuxing St, Taipei, 11031, Taiwan.
Biosens Bioelectron. 2022 Jan 1;195:113672. doi: 10.1016/j.bios.2021.113672. Epub 2021 Sep 28.
We present the first combination of a microfluidic polymerase chain reaction (PCR) with a gold nanoslit-based surface plasmon resonance (SPR) sensor for detecting the DNA sequence of latent membrane protein 1 (LMP1). The PCR microchannel was produced through a laser scribing technique, and the SPR nanoslit chip was manufactured via hot-embossing nanoimprinting lithography. Afterward, the LMP1 DNA probe was adsorbed onto the SPR chip of the integrated device through electrostatic interactions for further detection. The device can complete the analytical procedure in around 36 min, while the traditional machine requires 105 min to achieve similar signals under the same PCR thermal cycles. The calibration curve with serially diluted LMP1 DNA exhibited the accuracy (R > 0.99) and sensitivity (limit of detection: ∼10 g/mL) of the device. Moreover, extracted DNA from Epstein-Barr virus (EBV)-positive cells were directly detected through the integrated chip. In brief, this all-in-one chip can amplify gene fragments at the front-end and detect them at the back-end, decreasing the time required for the analysis without compromising accuracy or sensitivity. We believe this label-free, real-time, low-cost device has enormous potential for rapid detection of various viruses, such as EBV and COVID-19.
我们首次将微流控聚合酶链反应（PCR）与基于金纳米缝的表面等离子体共振（SPR）传感器相结合，用于检测潜伏膜蛋白 1（LMP1）的 DNA 序列。PCR 微通道通过激光划线技术制作，SPR 纳米缝芯片通过热压印纳米压印光刻技术制造。然后，通过静电相互作用将 LMP1 DNA 探针吸附到集成器件的 SPR 芯片上进行进一步检测。该设备大约可以在 36 分钟内完成分析过程，而传统的机器在相同的 PCR 热循环下需要 105 分钟才能达到类似的信号。用连续稀释的 LMP1 DNA 制作的校准曲线显示了该设备的准确性（R>0.99）和灵敏度（检测限：∼10 g/mL）。此外，通过集成芯片可以直接检测 Epstein-Barr 病毒（EBV）阳性细胞中的提取 DNA。总之，这种一体化芯片可以在前端放大基因片段，并在后端进行检测，在不影响准确性或灵敏度的情况下减少分析所需的时间。我们相信，这种无标记、实时、低成本的设备具有巨大的潜力，可用于快速检测各种病毒，如 EBV 和 COVID-19。
