應用粒子布朗運動檢測技術結合漸逝波照明於血清中量測C反應蛋白濃度

Abstract

近年來已有許多醫學報告指出血清中C反應蛋白的濃度可以作為心血管疾病危險因子指標，在本研究中，吾人藉由量測血清中螢光抗體粒子球之布朗運動做為檢測機制去偵測血清中Ｃ反應蛋白的濃度，當血清中之Ｃ反應蛋白抗原與螢光粒子表面之抗體反應時，粒子粒徑會變大，而造成其布朗運動速度值逐漸減慢，吾人在本研究中使用全反射螢光顯微技術以及微粒子追蹤測速技術分析得到抗原與抗體粒子球之結合過程中的布朗擴散特徵，在本研究中吾人成功量測十二種抗原濃度之血清，並利用此結果將心血管疾病之低、中、高危險指標區域中做更精細的分級，而由實驗結果可以看出在抗原抗體結合過程中其螢光粒子粒徑變化程度與血清中之抗原濃度相關。在高濃度抗原血清中，因為相同體積下抗原數目較多因而使抗體粒子球粒徑變化較大，進而造成其速度變化趨勢較低濃度抗原血清中之螢光粒子快。 在未來可以使用這機制簡單的檢測技術大量收集不同血清樣本的速度時間曲線資料，並且使每一個速度曲線都相對應血清樣本內某特定抗原濃度，已期待可以實際運用在心血管疾病危險因子之臨床檢測上。

The concentration of C-reactive protein (CRP) in serum is the symptom of risk factor for cardiovascular diseases (CVD). In this research, a new bio-sensing technique was applied to detect the concentration of CRP in serum by measuring the Brownian motion of fluorescent particles whose surface has been coated with anti-CRP. When CRP antigens react with anti-CRP coated on the surface of fluorescent particles, the diameter of particles will increase and the Brownian motion velocity of particle gradually slows down. Both Total Internal Reflection Fluorescence Microscopy (TIRFM) and Micro-Particle-Tracking-Velocimetry (μ-PTV) technique are utilized to obtain the characteristics of particles’ Brownian diffusion during the reaction process. In this study, I totally measure twelve different samples with different CRP concentration, and risk region for cardiovascular disease are successfully divided into twelve regions by different Brownian velocity profiles in different concentrations of CRP. The experiment results show that the degree of variation in beads’ diameters relates to the concentration of CRP antigen in serum during the binding process. Because there are larger amount of antigen in serum with higher concentration of CRP antigen, it leads to Brownian velocity of the particles in higher antigen concentration change more rapidly than Brownian velocity of the particles in lower antigen concentration. In the future, this simple sensing technique can be used to collect a large of Brownian velocity profile data, and every profile can represent each situation of specific antigen concentration and be applied to the clinical assay of cardiovascular diseases risk factors.