菌血症患者若不盡早治療，可能造成急性多器官轉移性感染，具有高危險性。因此在感染初期必須立即分辨是真菌、革蘭氏陽性菌或革蘭氏陰性菌，可縮小藥物選擇範圍而有效治療，並可避免因錯用抗生素而造成抗藥性。古生嗜鹽菌Halobacterium salinarum之紫色細胞膜 (purple membrane, PM) 含有細菌視紫質(bacteriorhodopsin, BR)，為一種光驅動質子泵浦，被光照射後可將質子由PM膜內側推向外側，造成質子梯度而產生光電流。利用PM膜光電流與入射光強度呈正相關，以及利用菌體本身可散射光之特性，我們先前已製備出一系列以PM為基礎之微生物光電晶片。本論文分為四部分，第一部分製作可捕捉真菌之核酸適體-PM複合晶片上，並探討晶片製程與最適化檢測條件。第二部分為使用可辨認脂磷壁酸 (lipoteichoic acid) 的抗體，進行可捕捉革蘭氏陽性菌之抗體-PM複合晶片之開發。第三部分則利用可辨認脂多醣 (lipopolysaccharide) 之抗體，開發可檢測革蘭氏陰性菌之免疫檢測晶片。最後，則以這三種晶片針對台灣常見之七種菌血症菌株進行檢測，結果發現，所開發晶片均可有效區分菌種類別。本研究所新開發的微生物感測晶片具有直接檢測、快速與高靈敏度之諸多優點，未來可應用於快速篩檢。

Untreated bacteremia patients are at high risk of developing acute multiple-organ metastatic infection. Early identification of infectious microorganisms as fungus, gram-negative bacteria, or gram-positive bacteria help narrow the antibiotics treatment, and avoid the development of antibiotic resistance. Halobacterium salinarum purple membrane (PM) contain bacteriorhodopsin, a light-driven proton pump able to transport protons across PM upon illumination and subsequently generate photocurrents. According to the linear dependence of PM photocurrents on illumination intensities, we had developed a series of different PM-based biosensors. This thesis comprises four parts. First, a PM chip coated with chitin-aptamers to detect fungi with optimization investigation. Secondly, antibodies against lipoteichoic acid (LTA) were immobilized on a PM chip to capture gram-positive bacteria. Thirdly, another immunosensing PM-based chip was constructed with antibodies against lipopolysaccharide (LPS) to recognize gram-negative bacteria. Finally, the performance of those three chips were investigated with seven bacteremia strains most commonly prevalent in Taiwan, demonstrating their differential recognition ability among fungi, gram-negative bacteria, and gram-positive bacteria. The newly developed PM-based bacteremia biosensor has the advantages of direct and fast detection with high sensitivity, and can be applied to rapid screening in the future.

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