臺灣博碩士論文加值系統

目前臨床上檢測細菌種類及抗藥性的方法，皆需要培養數天，過程耗時，故需要開發快速檢測方式。本研究以表面增強拉曼光譜對革蘭氏陰性菌嗜麥芽寡養單胞菌進行分析。將待測菌液滴上奈米銀粒子基板，產生的電漿現象增強拉曼訊號，即為表面增強拉曼光譜，可增加檢測的敏感性。本研究挑選出對抗生素氯黴素、匹美西林不同抗藥性表現的3株 S. maltophilia 菌株做培養，並以定時的方式做拉曼光譜的監測，藉由奈米銀基板所激發的表面增強拉曼效應，增強拉曼系統的偵測敏感度來分析菌株是否被抗生素殺死，進而達到快速檢測細菌抗藥性的目的。實驗結果可知，細菌與抗生素培養數小時後，偵測到細菌體內胺基酸或核酸的拉曼峰值訊號，表示細菌被抗生素所破壞，此樣本即為藥物敏感細菌。拉曼光譜系統利用奈米銀基板的表面增強拉曼訊號技術，來檢測細菌抗藥性，相較於傳統培養檢驗方式需要24小時時間，從實驗中我們成功縮短成在數小時內即可偵測細菌是否具有抗藥性。

In clinical, the traditional diagnosis method of the drug-resistant bacteria are time-consuming culture processes. This study utilizes surface enhanced Raman scattering (SERS) to get a rapid diagnosis of drug-resistant bacterial. We chose three different S. maltophilia strains, KJ, KJ2, and KJ2ΔTolC, and culturing with different type of antibiotics, chloramphenicol and piperacillin. Then, put the bacterial sample on the Ag nanoparticle substrate. The Ag nanoparticle will excited the plasmons phenomenon to enhanced Raman signal, it can increase the sensitivity of detection.From the result, it can detect drug-susceptible bacteria from Raman spectra after 6 hrs culturing. Compare to multidrug resistance strains, the different Raman peaks are the component inside of bacteria, such as amino acid, nucleic acid. It means that the antibiotics destroyed the drug-susceptible strains, and Ag substrate enhanced Raman signal of the outflow of the bacteria. Detection of drug-susceptible bacteria by using Ag nanoparticle SERS substrate, we can shorten the culture time to 6 hours. Contrast to traditional method, Raman spectroscopy with nanoparticle substrate is more rapidly detection the multidrug resistance in S. maltophilia.

目錄
誌謝 i
Abstract ii
中文摘要 iii
圖目錄 vi
表目錄 vii
第一章 序論 1
1.1 研究動機 1
1.2 研究背景 3
1.2.1 嗜麥芽寡養單胞菌(Stenotrophomonas maltophilia) 3
1.2.2抗生素種類機制介紹 7
1.2.3抗藥機制 11
1.2.4現行細菌抗藥性檢驗法 12
1.2 研究目的與文獻回顧 13
1.2.1拉曼光譜以及表面增強拉曼光譜的在細菌診斷上的應用 13
1.2.2相關文獻回顧 13
第二章 基礎理論 14
2.1 拉曼光譜 14
2.2 表面增強拉曼光譜 17
2.2.1 簡史 17
2.3拉曼光譜儀架設 21
第三章 材料與方法 22
3.1 實驗儀器介紹 22
3.2細菌樣本 24
3.3實驗材料及藥品 25
3.4 樣本前處理流程 25
3.4 光譜量測與處理 26
3.4.1 光譜處理 26
3.4.2 光譜分析 27
第四章 實驗結果 28
4.1奈米銀基板品質 28
4.2 S. maltophilia拉曼光譜 30
4.3與抗生素培養之拉曼光譜比較 31
4.4計算拉曼光譜峰值的比值732 cm-1/655 cm-1 38
4.5 S. maltophilia電顯圖 40
參考文獻 41

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