基於共振角度位移量測之表面電漿子共振生物感測器應用於抗青霉素金黃色葡萄球菌的恆溫擴增即時偵測

Abstract

生物感測器是近年來研究的熱門領域，表面電漿子共振應用於生物感測對生物分子進行偵測，由於實驗操作相對複雜，受限於操作人員培訓之繁瑣，無法真正普及商業用途。同時生物檢測方法不斷的發展，根據不同的檢測需求可以找到不同的檢測方法進行優化，但生物檢測往往受限於檢測時間較長且操作繁瑣，對於日益擴大的病患需求，加快檢測速度的方式顯得更為急迫。 本研究提出以共振角度位移量測的表面電漿子共振與LAMP生物檢測方法相結合以加快檢測之速率。本論文對表面電漿子共振原理進行研究應用，嘗試利用LAMP檢測核酸分子的方式與表面電漿子共振結合，提出SPR-LAMP的機構，並以此機構應用於抗甲氧西林金黃色葡萄球菌的檢測。本研究從機構設計和生物設計兩方面進行介紹，並對二者結合所作之機構改進，設計了一系列的實驗對系統性能進行驗證。本研究從機構設計和生物設計兩方面進行介紹，並對二者結合所作之機構改進，設計了一系列的實驗對系統性能進行驗證。該系統架構具有較好的穩定性，通過實驗量測系統之角度解析度為0.0154°，對應系統之偵測極限為1.23×10-4 RIU，敏感度為125°/RIU。LAMP於SPR機台之訊號可大致在15 min判定結果。針對實驗結果，對存在污染的原因與解決方式進行討論並提出了未來改進之方向。

Surface Plasmon Resonance (SPR) biosensor is a widely used from biological studies to clinical diagnosis application. In the situation of growing needs for real time detection, the SPR system has high potential in the early diagnosis. We proposed a SPR system based on angle shift which is more stable and easier to be used when different samples were tested. The SPR system integrated with isothermal amplification can be used in real time detection for DNA amplification. The mechanism of SPR was discussed and the design of our system is well explained in this thesis. Also, we made an intensive theoretical study of Loop mediated isothermal amplification (LAMP) and attempted to conduct the LAMP on SPR. Finally, we adapted the SPR system to amplify the gene mecA which is the marker of Methicillin-Resistant Staphylococcus Aureus (MRSA) within an hour. The angular resolution of our system is 0.0154°, the limit of detection is 1.23×10-4 RIU and the sensitivity is 125°/RIU. The signal of the LAMP can be detected by our SPR system. According to the problems showed during the experiment, reasons were discussed, conclusions were summarized and so are the future work.