Graduation Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei 10617, Taiwan.
Department of Electrical Engineering, National Taiwan University, Taipei 10617, Taiwan.
Biosensors (Basel). 2022 Dec 13;12(12):1163. doi: 10.3390/bios12121163.
Blood testing is a clinical diagnostic tool to evaluate physiological conditions, the immune system response, or the presence of infection from whole blood samples. Although conventional blood testing can provide rich biological information, it usually requires complicated and tedious whole blood processing steps operated by benchtop instruments and well-experienced technicians, limiting its usage in point-of-care (POC) settings. To address the above problems, we propose a microfluidic platform for on-chip plasma extraction directly from whole blood and in situ biomarker detection. Herein, we chose C-reactive protein (CRP) as the target biomarker, which can be used to predict fatal cardiovascular disease (CVD) events such as heart attacks and strokes. To achieve a rapid, undiluted, and high-purity on-chip plasma extraction, we combined two whole blood processing methods: (1) anti-D immunoglobulin-assisted sedimentation, and (2) membrane filtration. To perform in situ CRP detection, we fabricated a three-dimensional (3D) microchannel with an embedded electrochemical (EC) sensor, which has a modular design to attach the blood collector and buffer reservoir with standard Luer connectors. As a proof of concept, we first confirmed that the dual plasma extraction design achieved the same purity level as the standard centrifugation method with smaller sample (100 µL of plasma extracted from 400 µL of whole blood) and time (7 min) requirements. Next, we validated the functionalization protocol of the EC sensor, followed by evaluating the detection of CRP spiked in plasma and whole blood. Our microfluidic platform performed on-chip plasma extraction directly from whole blood and in situ CRP detection at a 0.1-10 μgmL concentration range, covering the CVD risk evaluation level of the high-sensitivity CRP (hs-CRP) test. Based on the above features, we believe that this platform constitutes a flexible way to integrate the processing of complex samples with accurate biomarker detection in a sample-to-answer POC platform, which can be applied in CVD risk monitoring under critical clinical situations.
血液检测是一种评估生理状况、免疫系统反应或全血样本感染的临床诊断工具。虽然常规血液检测可以提供丰富的生物学信息，但通常需要由台式仪器和经验丰富的技术人员操作复杂而繁琐的全血处理步骤，限制了其在即时检测（POC）环境中的应用。为了解决上述问题，我们提出了一种从全血中直接提取芯片血浆并进行原位生物标志物检测的微流控平台。在此，我们选择 C 反应蛋白（CRP）作为靶标生物标志物，它可用于预测致命性心血管疾病（CVD）事件，如心脏病发作和中风。为了实现快速、未稀释和高纯度的芯片血浆提取，我们结合了两种全血处理方法：（1）抗-D 免疫球蛋白辅助沉淀，和（2）膜过滤。为了进行原位 CRP 检测，我们制造了一个具有嵌入式电化学（EC）传感器的三维（3D）微通道，其具有模块化设计，可使用标准鲁尔接头连接血液收集器和缓冲储液器。作为概念验证，我们首先证实，双血浆提取设计以更小的样本（从 400 µL 全血中提取 100 µL 血浆）和时间（7 分钟）需求实现了与标准离心法相同的纯度水平。接下来，我们验证了 EC 传感器的功能化方案，随后评估了 CRP 在血浆和全血中的检测。我们的微流控平台直接从全血中进行芯片血浆提取和原位 CRP 检测，检测范围为 0.1-10 µgmL，涵盖了高敏 CRP（hs-CRP）检测的 CVD 风险评估水平。基于上述特点，我们相信该平台构成了一种灵活的方式，可以将复杂样品的处理与准确的生物标志物检测集成在一个即取即测的 POC 平台中，可应用于关键临床情况下的 CVD 风险监测。
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