無動力式微流道晶片進行全血血漿分離及原位比色法生物標記物檢測

Abstract

全血分析提供許多判斷人體健康狀況的參數，如各類血球數目、特定蛋白質濃度等，因此可以提供適當的治療方法。然而，傳統上通常需要訓練有素的醫學檢查人員，大型的量測儀器以及許多冗長的樣本處理過程來從全血樣本中提取血漿來進行血液分析。為了解決這些問題，我們希望研發一個整合比色分析的試紙與無須動力之真空式微流道系統進行微升體積的全血處理和分析。該系統共分為三個區域：一、血漿分離區域，二、檢測區域，三、吸入腔。對於血漿分離，我們設計了一個用於紅細胞沉降的三角形溝槽；同時，我們在裝置中加入了紅血球凝集因子(Anti-D)，以加快紅細胞沉降速率（erythrocyte sedimentation rate, ESR）。對於大多數臨床樣品，此晶片可在7分鐘內將未經過任何血液樣本處理步驟的50微升全血樣本中提取10微升的血漿。在血球沉降及血漿純化的過程中，紅血球的去除率約為99.99%，且未觀察到溶血作用。經由血液處理後的血漿將會自動流向比色試紙所在的檢測區域。本論文使用兩種比色試紙，分別為單純葡萄糖試紙檢測及整合奈米金粒之橫向流動免疫測定（Lateral flow immunoassay, LFIA）試紙進行C-reactive protein (CRP) 濃度檢測。上述測試皆可於20分鐘內完成，由於檢測的快速性和系統的便攜性，我們認為此設備具有發展成為定點照護（POC）系統的潛力。

Whole blood analysis is one of the most standard methods to understand or monitor one’s health conditions. To achieve a precise whole blood analysis results, it requires well-trained medical examiners, bulky instruments, and multiple sample pretreatment processes to extract blood plasma for blood analysis. To address these problems, we propose a vacuum-driven power-free microfluidic device integrated colorimetric test strip integrated for whole blood processing and analysis. The system consists of three regions: (1) the plasma separation region, (2) the detection region, and (3) the suction chamber. For plasma separation, we design a triangular trench for red blood cells (RBCs) sedimentation. Also, RBCs agglutination reagent was deposited on the trench to accelerate the erythrocyte sedimentation rate (ESR). For most clinical samples, our device can extract about 10 µL plasma from 50 μL whole blood without any sample pretreatment in 7 minutes. The RBCs removal rate is about 99.99% and no hemolysis effect is observed during the process. As for colorimetric detection, the extracted blood plasma automatically flows toward the detection region where a colorimetric-based test strip is located. In this thesis, we use two colorimetric test strips for demonstration, which are glucose colorimetric test strips for blood glucose measurement and gold nanoparticles integrated lateral flow immunoassay (LFIA) test strips for C-reactive protein (CRP) detection. All tests can be done within 20 minutes. Based on the above features, our device shows great potential to be applied in point-of-care (POC) applications.