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研究生(外文):Shu-Fan Tang
論文名稱(外文):A novel electrochemical method for the detection of Escherichia coli in liquid samples.
指導教授(外文):Ku-Shang ChangChung-Saint Lin
外文關鍵詞:amperometric sensorEscherichia colimethylene bluedetection of water quality
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大腸桿菌(Escherichia coli)為水質指標菌,因此有必要做此類水質大腸桿菌的監控;傳統檢測大腸桿菌群之方法常用為最確數法(most probable number, MPN),且需經過確定試驗(confirmed test)後才可鑑定大腸桿菌,主要係利用選擇性培養基來篩選微生物,有程序繁瑣費時且耗費成本等問題。
本研究結合安培電化學法及選擇性培養基來建立一套快速、簡便的檢測系統,應用選擇性培養基來篩選微生物,利用微生物代謝造成培養基電化學活性物改變造成之電化學反應變化來檢測微生物菌數,並篩選添加適當之電化學活性化學物質來放大訊號,結果顯示此系統對大腸桿菌鑑別度高,對E.coli之應答為同濃度Enterobacter等大腸桿菌群之30倍,可快速選擇性檢測大腸桿菌,檢測範圍為10 2 - 10 8 CFU/mL,線性關係良好(y = -0.8917x + 8.9141;R² = 0.9156),此檢測方法可用於大腸桿菌之快速檢測。

The typical coliform testing method, MPN, is time-consuming at 48 hours per sample. In addition, further analysis is required to differentiate fecal coliforms from E. coli in samples. A simple and rapid technique based on electrochemical principles is developed to analyze E. coli in liquid samples. The method includes a laboratory-built potentiostat connected with a pair of electrodes and a water sample with media containing lauryl sulfate. The electrodes were immersed in the liquid solution. When a water sample was placed into contact with the electrodes, the current change observed was used to determine the concentration of E. coli. If the current change exceeded a certain threshold, it indicated that the test sample contained E. coli. If the current change exerted to a low extent, or not at all, it was determined that the sample maybe contained other microorganisms (e.g., Enterobacter, Citrobacter, or Klebsiella). The sensor was able to acquire sufficient data within four hours at initial concentrations of E. coli higher than 105 CFU/ mL, making it suitable for use in real-time monitoring of bacterial growth. This system has potential application in the detection of E. coli concentrations in water of various sources.
謝誌 I
中文摘要 III
英文摘要 IV
目錄 VI
圖目錄 VIII
表目錄 X
第一章 前言 1
第二章 文獻整理 4
2.1大腸桿菌群之簡介 4
2.2大腸桿菌檢測方法簡介 5
2.3生物感測器(Biosensor) 9
2.4以電化學生物感測器開發多功能檢測平台之潛力 10
2.5以電化學感測器檢測大腸桿菌之目前研究結果 11
第三章 材料與方法 13
3.1 儀器設備 13
3.2 培養基 14
3.3 化學試藥 14
3.4 緩衝溶液 15
3.5 試驗用菌株 15
3.6電化學檢測系統之架構 19
3.6.1系統流程圖 20
3.7電化學檢測機制 23
3.8循環掃描伏安法(Cyclic voltammetry, CV) 30
3.9 API 10 S與API 20 E微生物生化鑑定套組 30
3.10原子力顯微鏡(Atomic Force Microscopy, AFM) 觀測 31
3.11 SDS-PAGE電泳分析 33
第四章 結果與討論 36
4.1 檢測系統之最適電壓探討 41
4.2 pH 對微生物生長的影響 45
4.3 檢測系統之培養基探討 46
4.4 電化學系統檢測大腸桿菌之培養基碳源最佳化探討 52
4.5 電化學系統對不同微生物及實際水樣檢測之討論 57
4.6 以循環伏安法檢測細菌之生長 58
4.7 原子力顯微鏡(Atomic force microscopy, AFM)觀察菌體 66
4.8 SDS-PAGE分析 68
第五章 結論與未來展望 76
參考文獻 77
附錄 85

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