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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 陳建甫(Chien-Fu Chen) | |
| dc.contributor.author | Zhi-Shun Dong | en |
| dc.contributor.author | 董至舜 | zh_TW |
| dc.date.accessioned | 2021-06-08T03:30:53Z | - |
| dc.date.copyright | 2019-08-16 | |
| dc.date.issued | 2019 | |
| dc.date.submitted | 2019-08-13 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/21314 | - |
| dc.description.abstract | 本研究使用graphene quantum dots (GQDs) 與gold nanoclusters (AuNCs) 應用於金黃色葡萄球菌與綠膿桿菌之ELISA檢測在結合紙張平臺上，並結合可攜式螢光偵測儀整合出多工螢光細菌感染檢測系統。在兩種不同細菌的ELISA檢測中使用不同放射波長之螢光奈米材料，作為不同目標物辨識依據，其中GQDs用於金黃色葡萄球菌之protein A偵測，而AuNCs用於綠膿桿菌之exotoxin A之偵測。藉由螢光強度去定量目標物濃度，最後利用光偵測晶片結合Arduino UNO所開發之可攜式螢光偵測儀對檢測結果作分析與數據輸出。達到定點照護多工檢測目的。本研究對於奈米材料結合在ELISA於紙張平臺上進行偵測抗體與捕捉抗體最佳進行濃度之優化。將光偵測晶片做波長判讀與光強度感應測試，並且將光偵測晶片與Arduino UNO板結合3D列印外殼，使螢光偵測區域能夠達到仿暗室之效果，於正常光環境中也能偵測紙張平臺上兩種奈米材料之螢光。本研究之多工螢光細菌感染檢測系統，可以於30分鐘內完成細菌感染偵測。針對金黃色葡萄球菌之protein A與綠膿桿菌之exotoxin A之最低偵測濃度皆為0.1 ng/mL。其中protein A之結果是由螢光偵測裝置所輸出之B值作數據處理所得，而exotoxin A之結果是由R值作數據處理所得。本系統可以在同一時間內偵測兩種細菌感染。期望未來能結合更多種不同放射波長之奈米材料，對於更多細菌之生物標記物進行多工檢測。 | zh_TW |
| dc.description.abstract | In this study, we use graphene quantum dots (GQDs) and gold nanoclusters (AuNCs) for enzyme-linked immunosorbent assay (ELISA) detection of Staphylococcus aureus and Pseudomonas aeruginosa on a paper platform combined with a portable fluorescence detector to integrate multiplex fluorescence bacterial infection detection system. Fluorescent nanomaterials with different emission wavelengths were used in the paper-based ELISA as detection signals of two different bacteria. GQDs and AuNCs were used as the signal label for protein A (detection of Staphylococcus aureus) and exotoxin (detection of Pseudomonas aeruginosa), respectively. The fluorescence intensity is the quantification of the target concentration. After paper-based ELISA tests, the fluorescence results were detected by the light sensor chip and the data were analyzed by Arduino UNO based portable fluorescence detection system. We also optimize the concentrations of the detection antibody with nanomaterials and the capture antibody. In order to eliminate the environmental interference, the light sensor chip and the Arduino UNO board were combined with the 3D printing shell as a dark room for sensing. As a result, the limit of quantification (LOQ) values for Staphylococcus aureus protein A and Pseudomonas aeruginosa exotoxin A detections are both 0.1 ng/mL, and the multiplex bacterial detection can be finished in 30 minutes. It can be expected that our system can combine with other nanomaterials with different emission wavelengths to perform on-site multiplex biomarkers detection in a rapid and efficient way. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T03:30:53Z (GMT). No. of bitstreams: 1 ntu-108-R06543055-1.pdf: 2321813 bytes, checksum: c2e52cc4deb4ccdffd9b5701df91160a (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 論文口試委員會審定書 i
致謝 ii 中文摘要 iii Abstract iv 圖目錄 viii 表目錄 ix 第一章 前言與文獻回顧 1 1.1 兩種高威脅性之細菌感染 1 1.2 金黃色葡萄球菌 1 1.3 綠膿桿菌 1 1.4 傳統細菌感染檢測方式 2 1.5 以生化感測機制檢測細菌感染 2 1.5.1 電化學 2 1.5.2 噬菌體檢測細菌感染 4 1.5.3 FRET檢測細菌感染 5 1.6 本研究所開發之系統 6 第二章 實驗設計與流程 8 2.1 實驗藥劑 8 2.2 儀器設備 8 2.3 GQDs合成 8 2.4 AuNCs合成 9 2.5 修飾biotin於AuNCs與GQDs表面上 9 2.6 修飾Streptavidin於protein A antibody 與toxR antibody上 9 2.7 奈米材料與抗體之接合 10 2.8 ELISA紙張平臺製備 10 2.9 螢光偵測儀組裝 10 2.10 Protein A ELISA實驗步驟 12 2.11 Exotoxin A ELISA實驗步驟 14 第三章 實驗結果與討論 16 3.1 GQDs與AuNC之材料鑑定 17 3.2 GQDs修飾biotin最佳條件探討 19 3.3 ELISA三明治法最佳捕捉抗體濃度 21 3.4 ELISA三明治法最佳偵測抗體濃度 24 3.5 Exotoxin A ELISA之清洗方式測試 27 3.6 螢光偵測儀之光波長分辨可行性與裝置仿暗室效果 29 3.7 Protein A 與exotoxin A ELISA實驗結果之螢光訊號偵測 31 第四章 結論與展望 34 參考文獻 35 | |
| dc.language.iso | zh-TW | |
| dc.title | 攜帶式多工螢光細菌感染檢測系統之開發 | zh_TW |
| dc.title | Portable Fluorescent Detector for Multi-Target Bacterial Infection Diagnosis on a Paper-Based Device | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 107-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 葉怡均(Yi-Chun Yeh),余政儒(Cheng-Ju Yu) | |
| dc.subject.keyword | 石墨稀量子點,奈米金團簇,ELISA,紙張平臺,Arduino, | zh_TW |
| dc.subject.keyword | graphene quantum dots,gold nanoclusters,ELISA,paper-based platform,Arduino, | en |
| dc.relation.page | 37 | |
| dc.identifier.doi | 10.6342/NTU201902558 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2019-08-14 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 應用力學研究所 | zh_TW |
| Appears in Collections: | 應用力學研究所 | |
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| File | Size | Format | |
|---|---|---|---|
| ntu-108-1.pdf Restricted Access | 2.27 MB | Adobe PDF |
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