研究生: |
許敦淯 Tun-Yu Hsu |
---|---|
論文名稱: |
微生物生物光電感測晶片之開發及化粧品檢測之應用 Development of bio-photoelectric sensing chips for microorganisms and applications in cosmetic testing |
指導教授: |
陳秀美
Hsiu-Mei Chen |
口試委員: |
陳秀美
Hsiu-Mei Chen 戴守谷 Shou-Ku Tai 賴惠敏 Hui-Min Lai 廖國宸 Kuo-Chen Liao 葉旻鑫 Min-Hsin Yeh |
學位類別: |
碩士 Master |
系所名稱: |
工程學院 - 化學工程系 Department of Chemical Engineering |
論文出版年: | 2023 |
畢業學年度: | 111 |
語文別: | 中文 |
論文頁數: | 182 |
中文關鍵詞: | 生物光電感測晶片 、化粧品檢測 、大腸桿菌 、綠膿桿菌 、金黃色葡萄球菌 、白色念珠菌 |
外文關鍵詞: | Bio-photonic Sensor Chip, Cosmetic testing, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans |
相關次數: | 點閱:176 下載:0 |
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紫膜 (purple membrane, PM)為Halobacterium salinarum細胞膜成分之一,其內部具有細菌視紫質 ( bacteriorhodopsin, BR) 。BR具有高穩定性也是一個光驅動質子泵浦,透過BR光驅動質子泵產生可被偵測的電流訊號,可以作為生物光電傳感器的信號轉換器。
先前已經利用單層紫膜晶片開發出可檢測微生物的紫膜光電感應晶片,具有高檢測靈敏度還有快速檢測的特性。傳統檢測化粧品方法大都具有操作步驟繁瑣、檢測時間長、靈敏度低等缺點。國內外的化粧品規範中,大腸桿菌、綠膿桿菌、金黃色葡萄球菌、白色念珠菌此四株菌被列為不得檢出的菌。本文即針對這四株不得檢出菌開發出高特異性的各種抗體晶片,並進行針對單一菌種、混合菌液、化粧品樣品之分析。結果顯示,所開發出的四種抗體-PM生物光電感測晶片能各個別檢出其目標菌株,最低檢測濃度均為 1 Colony-forming unit (CFU)/mL,且各種抗體檢測晶片也都具有高專一性在混和菌液中可明顯分別目標菌。
對於不同劑型的化粧品(液劑、乳劑、霜劑)的檢體時,四種抗體-PM生物光電感測晶片檢測結果都與平板計數法得出的結果是一致的。化粧品防腐效能的測試中也顯示晶片的檢測結果是符合測試指引的。
最後利用結合奈米金與適體的複合粒子再去標定被抗體晶片所捕捉的菌株,結果發現可以使晶片的檢測靈敏度更加提升,使其最低檢測濃度可低達1 CFU/100 mL。
本研究所開發的四種微生物感測晶片具有直接且快速檢測、高靈敏度的優點,未來可以應用於食品、水質的檢測。
Purple membrane (PM) resides in the cellular membrane of Halobacterium salinarum and contains bacteriorhodopsin (BR). BR has high stability and functions as a light-driven proton pump, which drives photocurrent production, and hence can be applied as the signal transducer of a photoelectric biosensor. In our previous studies, a PM monolayer-coated electrode was devised and applied to detect microorganisms, with the advantages of high sensitivity and rapid detection. For cosmetics, plate counting is the traditional gold-standard way to detect microorganisms, suffering the disadvantages of complex and time-consuming procedures, low sensitivity, and requirement of professional operators. According to domestic and international cosmetic regulations, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans are considered must-not-be-detectable bacteria. In this thesis, four highly sensitive and specific PM-based microbial biosensors are developed for the detection of these four strains respectively, in single-strain, mixed-strain, and cosmetic samples. The results show that each biosensor detects its target strain with a limit of detection of 1 CFU/mL. All four biosensors have high specificity with the capability to distinctly discriminate their own target strain from the other three strains in a mixed solution. The results of microbial tests in liquid, emulsion, and cream forms of cosmetics using biosensors are consistent with those using the plate-count method. The test on the antiseptic efficacy of cosmetics demonstrates the applicability of those four biosensors. Finally, using gold-nanoparticle-labeled aptamers with sequences specific to the target strain, the sensitivity of each biosensor is further enhanced with a limit of detection of 1 CFU/100 mL. This research demonstrates the feasibility of the applicability of PM-based microbial biosensors in microbial detection of cosmetics.
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