使用適體修飾之延伸閘極場效應電晶體結合壓電微泵驅動微流體系統進行全血處理及心肌旋轉蛋白檢測

Abstract

急性心肌梗塞 (AMI) 是指冠狀動脈突然阻塞的情況。作為急性冠狀動脈綜合症 (ACS) 的一種嚴重形式，即時檢測AMI對於防止永久性心臟損傷和心衰竭至關重要。在臨床環境中，免疫分析是疑似心血管疾病（CVDs）生物標誌物分析的主要方法，使用的技術包括化學發光、螢光和酵素結合免疫吸附法（ELISA）。然而，這些方法面臨著非特異性吸附、高溫下有限的耐久性以及用於生物檢測中複雜的抗體修飾等困難。為了克服這些挑戰，我們建立了一種功能化適體的延伸式閘極場效電晶體（EGFET），並結合一個全血處理的微流體平台用於檢測心肌旋轉蛋白（cTnI）。透過與目標抗原結合並引起適體結構的構象變化來提高感測性能。我們的EGFET-適體感測器能夠在0.1 pg/mL到1000 pg/mL的廣泛動態範圍內顯示出良好的對數線性性能。此外，我們有效地使用了氯化銀銀膠（Ag/AgCl）作為參考電極，它可以方便地整合到我們的微流體平台中。我們也透過微流體系統控制樣本與檢測溶液的流動來改進了檢測過程，使操作更加簡單有序。延伸閘極場效應電晶體-適體感測器和微流體平台的結合減少了處理血液所需的時間，並能夠精確測量全血中的心肌旋轉蛋白，顯示出在臨床檢測應用中的巨大潛力。

Acute myocardial infarction (AMI) occurs when a coronary artery is suddenly blocked. As a severe form of acute coronary syndrome (ACS), timely detection of AMI is crucial to prevent permanent cardiac damage and heart failure. In clinical settings, immunoanalysis is the primary method for biomarker analysis in suspected cardiovascular diseases (CVDs), using techniques such as chemiluminescence, fluorescence, and ELISA. However, these methods face difficulties such as nonspecific adsorption, limited durability at high temperatures, and complex antibody modifications for biological detection. To overcome these challenges, we established an aptamer-functionalized Extended Gate Field-Effect Transistor (EGFET) integrated with a whole-blood processing microfluidic platform for troponin I (cTnI) detection. This decision improves sensing performance by increasing target binding and causing conformational changes in the aptamer structure. Our EGFET-aptasensor showed good log-linear performance over a broad dynamic range of 0.1 pg/mL to 1000 pg/mL. Additionally, we effectively used reference electrode silver chloride (Ag/AgCl) paste, which is conveniently incorporated into our microfluidic platform. We enhanced the process of detection for more straightforward and organized operations by managing the flow of samples and detection buffers. The combination of the EGFET-aptasensor and microfluidic platform reduced the time required for processing blood and allowed for precise measurement of cTnI spiked in whole blood, showing great promise for point-of-care applications.