1Department of Chemical and Biomolecular Engineering, University of Notre Dame, Notre Dame, IN 46556 USA.
2Center for Microfluidics and Medical Diagnostics, University of Notre Dame, Notre Dame, IN 46556 USA.
Commun Biol. 2019 May 20;2:189. doi: 10.1038/s42003-019-0435-1. eCollection 2019.
Extracellular vesicles (EV) containing microRNAs (miRNAs) have tremendous potential as biomarkers for the early detection of disease. Here, we present a simple and rapid PCR-free integrated microfluidics platform capable of absolute quantification (<10% uncertainty) of both free-floating miRNAs and EV-miRNAs in plasma with 1 pM detection sensitivity. The assay time is only 30 minutes as opposed to 13 h and requires only ~20 μL of sample as oppose to 1 mL for conventional RT-qPCR techniques. The platform integrates a surface acoustic wave (SAW) EV lysing microfluidic chip with a concentration and sensing microfluidic chip incorporating an electrokinetic membrane sensor that is based on non-equilibrium ionic currents. Unlike conventional RT-qPCR methods, this technology does not require EV extraction, RNA purification, reverse transcription, or amplification. This platform can be easily extended for other RNA and DNA targets of interest, thus providing a viable screening tool for early disease diagnosis, prognosis, and monitoring of therapeutic response.
细胞外囊泡 (EV) 中含有 microRNAs (miRNAs)，它们具有作为疾病早期检测生物标志物的巨大潜力。在这里，我们提出了一种简单、快速的无 PCR 集成微流控平台，能够绝对定量 (<10%的不确定性) 血浆中游离 miRNAs 和 EV-miRNAs，检测灵敏度为 1 pM。与传统的 RT-qPCR 技术相比，该测定时间仅为 30 分钟，而不是 13 小时，所需样本量仅为 ~20 μL，而不是 1 mL。该平台集成了表面声波 (SAW) EV 裂解微流控芯片与浓度和传感微流控芯片，该芯片采用基于非平衡离子电流的电动膜传感器。与传统的 RT-qPCR 方法不同，该技术不需要 EV 提取、RNA 纯化、逆转录或扩增。该平台可以很容易地扩展到其他感兴趣的 RNA 和 DNA 靶标，从而为早期疾病诊断、预后和治疗反应监测提供了一种可行的筛选工具。
