Simultaneous Electrochemical Detection of miRNAs in Human Body Fluids: Development of an Integrated Multiplexed Microfluidic RNA sensor

本项目旨在开发一种快速，可靠和超灵敏的电化学MicroRNA（miRNA）芯片实验室（lab-on-a-chip，简称miRNA）检测系统，该系统具有多路微流体技术，可以早期，轻松和同时测量临床相关样本（包括但不限于血浆，唾液，尿液，血清甚至全血）中的不同miRNA。此外，它还致力于发现不同的miRNAs作为肺肿瘤和间质性肺病（ILD）的潜在生物标志物，这些miRNAs可以用于临床诊断和预后目的，包括在这种基于芯片的微流控平台中，早期发现致病因子在临床医学中起着关键作用，将增加患者的生存机会。MiRNA是一种小RNA分子，在基因表达的调节中发挥着关键的调节作用。一些miRNAs的表达具有疾病特异性，其表达浓度的增加或减少都可以被检测到，因此，miRNAs作为肿瘤、心血管疾病和间质性肺病的生物标志物越来越重要。在过去的十年中，已经证明miRNA在临床相关样品（例如血浆、血清、唾液和尿液）中保存良好，并且显示出比基于蛋白质的生物标志物高得多的灵敏度。这些方面将有利于基于miRNA的检测系统作为未来诊断设备的标准，并将允许我们使用其他样品，如尿液或唾液和/或使用低样品体积进行检测。小型化的诊断系统设备提供了对各种生物标志物的特异性和灵敏度检测的加速，其可以适用于许多医学领域-从更便宜的临床试验，早期诊断和治疗复杂疾病。微流控技术和适于大规模生产的制造的组合允许构建低成本和便携式护理点（POC）诊断系统，这导致在速度和低样品消耗方面的额外优势。（微技术，微流体和生物医学）将有助于ILD的诊断和治疗指导。所提出的miRNA检测系统的成功演示将提供无需扩增即可直接测量miRNA的优势。原理测量的证明显示了非常有希望的结果，并且让我们激励将现有的免疫传感器与不同的扩增方法相结合，以实现无需昂贵的PCR扩增步骤的miRNA分析平台。

The present project aims to develop a rapid, reliable, and ultrasensitive electrochemical MicroRNA (miRNA) lab-on-a-chip (LOC) detection system with multiplexed microfluidics to gauge different miRNAs in clinically relevant samples (including but not limited to blood plasma, saliva, urine, serum or even whole blood) early, easy and simultaneously. Furthermore, it is dedicated to the discovery of different miRNAs as potential biomarkers for lung tumors and interstitial lung diseases (ILD), which can be employed later for diagnostic and prognostic purposes in the clinical settings, included in such a chip-based microfluidic platform.The early detection of disease-causing agents plays a key role in the clinical medicine and will increase the survival chances of the patients. MiRNAs are small RNA molecules that play a key regulatory role in the regulation of gene expression. Some miRNAs are expressed in a disease-specific manner and can be detected in either increased or decreased concentration.Therefore, miRNAs are gaining increasingly importance as biomarkers for cancer, cardiovascular disease and interstitial lung diseases. During the last decade, it has been demonstrated that miRNAs are well preserved in clinically relevant samples (e.g. blood plasma, serum, saliva and urine) and show considerably higher sensitivity than protein-based biomarkers. These aspects will favor miRNAs based detection systems as a standard in the future diagnostics devices and will allow us to use other samples like urine, or saliva and/or to work with low sample volumes for the detection.Miniaturized diagnostic system devices offer an acceleration of the specific and sensitive detection of various biomarkers, which can be adapted into many areas of medicine - from cheaper clinical trials, to early diagnosis and treatment of complex diseases. The combination of microfluidics and fabrication suitable for mass production allows for the construction of low-cost and portable point of care (POC) diagnostic systems which results in additional advantages in terms of speed, and low sample consumption.The new miRNA detection method realized by this proposal using a unique combination of different research areas (Microtechnology, microfluidics and biomedicine) will assist diagnosis and therapy guiding in ILD. The successful demonstration of the proposed miRNA detection system will provide the advantage that miRNA can be measured directly without amplification. The proof of principle measurements show very promising results and let us motivate to combine the existing immunosensors with different amplifications approaches to achieve a miRNA profiling platform without expensive PCR amplification step.