本研究之設計概念與實驗執行在於利用微生物晶片技術，結合銀奈米粒子並配合金屬銀析出之反應，且以光學式與電阻式偵測系統來量測免疫反應訊號，建立起新型之免疫分析系統，提供另一種有效且方便的免疫分析檢測方式。美國根據國家院內感染監測系統統計，院內感染病人身上培養出的菌株，金黃色葡萄球菌佔13%，金黃色葡萄球菌是造成嚴重感染菌血症之重要病原菌，而且近年來有逐漸增加之趨勢。所以在抗原抗體標定的選取上，利用金黃色葡萄球菌特有的protein A與免疫球蛋白G (IgG) 作為免疫分析模型，以探討新型免疫分析檢驗之可行性與效能。此外，在微晶片上執行免疫分析反應具有改良多樣性、低樣本用量與高靈敏度等特性，且可以縮短偵測反應的時間。實驗的架構朝向臨床上廣泛使用的三明治免疫分析模式，利用上下兩層已知抗體來對中間待測抗原作定性與定量之分析。有別於酵素連結免疫吸附分析(enzyme-linked immunosorbent assay, ELISA)，我們採用現行抗原抗體在晶片表面之固定技術，並加入銀析出增強溶液，來增顯免疫的反應偵測訊號。利用光學與電性式偵測結果顯示，本研究方法已可完成免疫分析的步驟與目標，光學與電性偵測靈敏度分別達到10-9 g/mL (1 ng/mL)與10-10 g/mL (0.1 ng/mL)，且使用的偵測方式也可搭配銀析出現象產生可供檢測之訊號，大幅縮短檢驗時間為30分鐘內，比傳統ELISA檢驗所需3到4小時，更為快速。此外，利用微機電技術、奈米粒子標記技術與銀訊號放大增強方法，建立一個全新、快速且靈敏之免疫分析系統，相信本研究之建立，未來將對臨床免疫分析方法提供嶄新的觀念與貢獻。

In this study, silver nanoparticle-labeled antibody is coupled with silver enhancement method and microchip to provide an effective and convenient immunoassay. Besides, the immunoassay has the advantages of versatile applications, lower sample consumption, high sensitivity and shorter detection time. According to the survey of infection system, Staphylococcus aureus is the main factor to lead to bacteremia. The application of immune analysis is widespread in biomedical field, because it is sensitivity and specificity. The design and experiment of this research lie in combining the biochip and nanoparticles to set up a new immunoassay system. In this study, Protein A from S. aureus and immunoglobulin G (IgG) are selected as the model immunoassay to estimate the feasibility and efficiency of the novel immunoassay. There is major format developed in this study, sandwich immunoassay (three-layer format), primary and secondary antibodies are used to quantify and qualify test antigens. Compared to traditional enzyme-linked immunosorbent assay (ELISA), we adopt the immobilization of antigen or antibody on the chip surface, and introduce silver enhancement method to amplify the detection signal generated by the silver nanoparticles. The experimental data of optical and electric detection show that the silver-precipitation immunoassay can be worked. The relationship between sample concentration and detection signal is discussed and the detection limit (sandwich assay) for the sample antigen are 10-9 g/mL (1 ng/mL) and 10-10 g/mL (0.1 ng/mL) in optical and electric system respectively. Further, the detection time of our system is 30 minutes shorter than 3 to 4 days of transitional ELISA method. In addition, we combine MEMS technology and silver enhancement to achieve a novel immunoassay system. The high applicability and biochemical efficiency of this study can provide an alternative for rapid, sensitive and convenient immunoassay.

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