參考電極嵌入雙微反應槽整合離子敏感場效應電晶體應用於細菌抗藥性測試

Abstract

敗血症是一種具有高死亡率的疾病，尤其是在老年人，免疫功能低下和患病的患者中。嚴重的創傷和某些醫療程序，例如靜脈輸血和裝置或器官的植入，很可能導致細菌性血液感染（BSI）並導致嚴重的敗血症。儘管醫療技術和環境已有飛速的發展，敗血症仍然是全世界的主要死亡原因，死亡率約為50％，早期使用有效的抗生素治療可以大大降低發病率和死亡率。傳統上若需要進行細菌藥敏測試通常需要一至兩天的時間，其中包含血瓶機培養放大血液中的細菌濃度以及繁瑣的細菌清洗流程，用以進行後續的細菌抗藥性檢測，上述冗長及繁複的人工流程不利於需要即時使用適當抗生素治療的病患。因此，如何能即時且快速的進行細菌的鑑定以及抗藥性測試是十分重要的。為了解決此問題，在研究中我們利用對酸鹼值十分敏感的離子感測場效電晶體(ion-sensitive field-effect transistor)與微流道技術，結合細菌生長時會消耗醣類並釋放有機酸的特點，將不同菌株小體積細菌液培養在晶片上，並且利用可攜式儀器進行三十分鐘的短時間細菌藥敏測試，我們目前已經成功在單一晶片上進行兩種菌株的藥敏測試，具有抗藥性的菌株確實生長情形會比沒抗藥性菌株還要顯著。期望在未來的研究上能增加晶片感測面積以及晶片靈敏度以完成更多細菌樣本量的快速檢測。

Sepsis is a common cause of morbidity and mortality, particularly in elderly, immunocompromised, and ill patients. Serious trauma and certain medical procedures, like intravenous transfusion and implantation of devices or organs, may cause bacterial bloodstream infections (BSI) or even lead to severe sepsis. An early treatment with effective antibiotics would greatly reduce the risk of morbidity and mortality. The dosage required for different bacteria will be different, and the method of finding most appropriate dosage depends on the antibiotic susceptibility test. Traditionally, it takes one to two days to perform an antibiotic susceptibility test, requiring a large incubator and tedious manual processes. In this thesis, we integrated dual microchamber with ion-sensitive field-effect transistor (ISFET) to perform real-time and dual bacteria growth monitoring. ISFET is very sensitive to pH value change of environment, which would happen when bacteria consume sugars and releasing organic acids during their growth phase. Here, we loaded two bacteria strains into microchamber and cultured on the sensor surface. Then, the ISFET sensor was connected to a portable instrument for 30-60 mins bacteria growth monitoring under various antibiotic treatment. Based on our results, the bacteria growth rate of the resistant strains is more significant than the susceptible strains, and we achieved rapid, label-free and real-time dual bacteria detection.