研究生: |
李俊葦 Lee, Chun-Wei |
---|---|
論文名稱: |
氧化還原活性金奈米粒子(raGNPs)在奈米篩分微流體系統(NS-MFS)中以電化學超敏感檢測菌血症並使用磺化聚苯胺作為兩性離子和導電界面進行電極上的抗生物污染 Ultra-Sensitive Electrochemical Detection of Bacteremia Enabled by Redox-Active Gold Nanoparticles (raGNPs) in a Nano-Sieving Microfluidic System (NS-MFS) with Sulfonated Polyaniline as Zwitterionic and Conductive Interfaces for Anti-biofouling on Electrodes |
指導教授: |
曾繁根
Tseng, Fan-Gang |
口試委員: |
張晃猷
Chang, Hwan-You 王本誠 Wang, Pen-Cheng 盧彥文 Lu, Yen-Wen 楊重熙 Yang, Chung-Hsi 范育睿 Fan, Yu-Jui |
學位類別: |
博士 Doctor |
系所名稱: |
原子科學院 - 工程與系統科學系 Department of Engineering and System Science |
論文出版年: | 2019 |
畢業學年度: | 107 |
語文別: | 中文 |
論文頁數: | 83 |
中文關鍵詞: | 電化學 、細菌 、聚苯胺 、抗生物汙染 |
外文關鍵詞: | Electrochemical, Bacteria, Polyaniline, Anti-biofouling |
相關次數: | 點閱:2 下載:0 |
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最標準的細菌檢測方式為利用培養基進行細菌增殖,再利用一系列生化反應,例如微生物發酵、特定受質的利用、還原與降解等特性來鑑定微生物種類,或是利用聚合酵素連鎖反應 (Polymerase Chain Reaction,PCR),質譜鑑定(Mass Spectrometry,MS)等大型機台進行判讀,這些方法精準度高,但至少需耗費一天以上作增殖反應,若遇上培養困難、生長緩慢或是無適當培養基的菌種再更為耗時。
本研究提出了一個奈米篩分微流體系統(Nano-Sieving Microfluidic System, NS-MFS)這是一個基於信號放大的檢測技術,該技術通過在氧化還原活性金奈米顆粒(redox active gold nanoparticles, raGNP)上結合電化學活性分子(electrochemical active molecule, EAM)和特異性抗體。經修飾的raGNP通過特異性抗體與細菌表面結合,再通過電化學聚合形成電化學活性的聚合物膜,當施加電壓時,EAM膜的氧化還原活性電流可以通過常規電化學檢測系統檢測,工作時間約30分鐘並達到~10 CFU/mL的生理檢測需求。
為了能夠在生醫檢測應用上,減少非特異性吸附等造成的背景雜訊干擾,在本研究中,我們提出將(self-doped sulfonated polyaniline, SPANI)作為塗層材料整合於Au電極上,該塗層材料在介電泳捕捉大腸桿菌與清洗的中顯示出優異的抗生物污染和導電特性,不僅具有非常低的殘留質量(1.438%),且在三循環實驗中捕捉細胞數量之標準差為22.47(PEG修飾表面的標準差高達208.9)顯示其具有高的電穩定性。未來應可實用在生物或醫學的快速檢測上。
The most standard method of bacterial detection is to use the culture medium for bacterial proliferation, and then use a series of biochemical reactions, such as microbial fermentation, specific substrate utilization, reduction and degradation, to identify microbial species, or to use large-scale machines such as polymerase chain reaction (PCR) or mass spectrometry (MS) for interpretation. These methods are highly accurate, but required at least one day for proliferative reactions. It is more time consuming to encounter strains that are difficult to grow, slow to grow, or have no suitable medium.
This study proposes a strategy based on signal amplification by conjugating an electrochemical active molecule (EAM) and a specific antibody on redox active gold nanoparticles (raGNPs). The modified raGNPs are conjugated to the surface of bacteria via a specific antibody to form an electrochemically active poly-film by electropolymerization. When applying a voltage, a redox-active current of the EAM film can be detected by a conventional electrochemical detection system. The working time is ~30 minutes and reached a physiological testing requirement of ~10 CFU/mL.
In order to reduce the background noise caused by non-specific adsorption in bio-detection applications, in this study, we propose to integrate self-doped sulfonated polyaniline (SPANI) as a coating material on the Au electrode. The coating material showed excellent bio-contamination and conductive properties during DEP capturing-and-releasing processes, not only with very low residual mass (1.438%), but also the standard deviation of the number of cells captured in the three-cycle experiment was 22.47. (The standard deviation of the PEG-modified surface is as high as 208.9) indicating its excellent electrical stability. The future should be practical for rapid detection of biology or medicine.
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