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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 張建成(Chien-Cheng Chang) | |
dc.contributor.author | Yuh-Shiuan Chien | en |
dc.contributor.author | 簡裕軒 | zh_TW |
dc.date.accessioned | 2021-06-07T23:44:39Z | - |
dc.date.copyright | 2020-08-20 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-08-14 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16733 | - |
dc.description.abstract | 本研究使用銅沉積訊號放大技術(Copper deposition-induced signal amplification) 應用於金黃色葡萄球菌與綠膿桿菌之側層流免疫測定法(Lateral Flow Imunoassay; LFIA)檢測,並結合可攜帶式光強度偵測儀整合出多工細菌檢測系統。其中本研究測定方法為使用側層流型態完成酵素結合免疫吸附分析法(Enzyme-linked immunosorbent assay; ELISA)反應,並且使用傳統金奈米粒子作為目標物辨識依據,側層流反應完成後以硫酸銅及抗壞血酸之還原反應進行訊號放大,用於金黃色葡萄球菌之Protein A偵測及用於綠膿桿菌之Exotoxin A之偵測。為了驗證該訊號放大方法,本研究利用量測光強度來定量目標物濃度,量測結果顯示具顯著放大效果,最後利用光強度偵測晶片結合Arduino UNO所開發之可攜帶式光強度偵測儀對檢測結果作分析與數據呈現。達到定點照護之多工檢測目的。本研究之多工細菌檢測系統,可以於25分鐘內完成細菌偵測。針對金黃色葡萄球菌之Protein A與綠膿桿菌之Exotoxin A進行訊號放大後其最低偵測濃度分別為3 ng/mL與10 ng/mL,與未放大前的偵測強度相差1個數量級。本系統可以在同一時間內偵測兩種細菌感染,並且相較於傳統快篩系統具有更好的靈敏性(sensitivity)。期望未來能結合更多種不同LFIA結構與更換不同放射波長的生物探針,對於更多細菌之生物標記物進行檢測,並擁有更好的檢測靈敏性與特異性。 | zh_TW |
dc.description.abstract | In this study, the copper deposition signal amplification technology was applied to the lateral laminar flow immunoassay detection of Staphylococcus aureus and Pseudomonas aeruginosa, and combined with a portable light intensity detector to integrate a multiplex bacteria detection system. The measurement method in this study is to use the lateral laminar flow pattern to complete the enzyme-binding immunosorbent analysis reaction, and use traditional gold nanoparticles as the basis for target identification. After the lateral laminar flow reaction is completed, the reduction reaction of copper sulfate and ascorbic acid is used to signal Zoom in for the detection of Protein A of Staphylococcus aureus and the detection of Exotoxin A of Pseudomonas aeruginosa. In order to verify the signal amplification method, this study uses the measured light intensity to quantify the concentration of the target. The measurement result shows a significant amplification effect. Finally, the light intensity detection chip combined with the portable light intensity detector developed by Arduino UNO Analyze the test results and present the data. To achieve the purpose of multi-work detection of designated care. The multi-tasking bacteria detection system in this study can complete bacteria detection within 25 minutes. After signal amplification of Protein A of Staphylococcus aureus and Exotoxin A of Pseudomonas aeruginosa, the lowest detection concentrations are 3 ng/mL and 10 ng/mL, respectively, which are an order of magnitude difference from the detection intensity before unamplification. This system can detect two bacterial infections at the same time, and has better sensitivity than traditional fast screening systems. It is hoped that in the future, it will be able to combine more different LFIA structures and replace bio-probes with different emission wavelengths to detect more bacterial biomarkers, and have better detection sensitivity and specificity. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T23:44:39Z (GMT). No. of bitstreams: 1 U0001-1008202014012300.pdf: 3546903 bytes, checksum: bef21d14afd22620b73a86b544bc8ec3 (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 論文口試委員審定書 i 致謝 ii 摘要 iii Abstract iv 目錄 vi 圖目錄 ix 表目錄 xi 第一章、前言與文獻回顧 1 1.1 細菌對公衛之影響 1 1.1.1金黃色葡萄球菌 1 1.1.2 綠膿桿菌 2 1.2 細菌檢測黃金標準 2 1.3 側層流免疫測定法 3 1.4以生化感測機制檢測細菌感染 4 1.4.1電化學 4 1.4.2 Cu-MOF奈米粒子檢測 6 1.4.3磁性奈米顆粒(Magnetic Nanoparticles;MNPs)修飾檢測 7 1.4.4 FRET檢測細菌感染 8 1.4.5拉曼散射(SERS)的側向流免疫法生物感測器 9 1.5 本研究所開發之系統 11 第二章、實驗原理 12 2.1 側層流 12 2.2 酵素結合免疫吸附分析法 12 2.3側層流免疫試片(Lateral Flow Immunoassay Strip) 14 第三章、材料與方法 18 3.1 實驗藥劑 18 3.2 儀器設備 18 3.3 樣品製備 19 3.3.1 Copper sulfate pentahydrate溶液配置 19 3.3.2 L-ascorbic acid (AA, reagent grade)溶液配置 19 3.3.3 Protein A抗原標準品配置 20 3.3.4 Exotoxin A抗原標準品配置 20 3.3.5 Protein A捕捉抗體配置 20 3.3.6 Exotoxin A捕捉抗體配置 20 3.3.7 Rabbit IgG捕捉抗體配置 21 3.3.8 奈米金接抗體製備 21 3.4 側層流免疫測定試片製備 21 3.5 光強度偵測儀製備 24 3.6 側層流免疫測定法實驗步驟 24 第四章、結果與討論 27 4.1 反應時間探討 29 4.2 NC membrane上捕捉抗體最佳化 31 4.3 Protein A與Exotoxin A抗原測試 34 4.4 Copper deposition-induced enhancement測試 36 4.5 多工檢測干擾實驗 40 4-6光強度檢測系統可行性測試 42 第五章、結論與未來展望 45 參考文獻 46 | |
dc.language.iso | zh-TW | |
dc.title | 利用銅沉積技術開發應用於多工細菌檢測之高靈敏度側層流免疫測定試紙元件 | zh_TW |
dc.title | Signal Amplified Gold Nanoparticles for Bacterial Infection Diagnosis on Lateral Flow Immunoassay Device | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 陳建甫(Chien-Fu Chen) | |
dc.contributor.oralexamcommittee | 朱錦洲(Chin-Chou Chu),林真真(Jen-Jen Lin),黃執中(Chih-Chung Huang),崔博翔(Po-Hsiang Tsui) | |
dc.subject.keyword | 銅沉積訊號放大,酵素結合免疫吸附分析法,側層流免疫測定法,金黃色葡萄球菌,綠膿桿菌,蛋白質A,外毒素A, | zh_TW |
dc.subject.keyword | Copper Deposition-induced Signal Amplify,Enzyme-linked immunosorbent assay,Lateral Flow Immunoassay Test,Staphylococcus aureus,Pseudomonas aeruginosa,Protein A,Exotoxin A, | en |
dc.relation.page | 49 | |
dc.identifier.doi | 10.6342/NTU202002797 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2020-08-16 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 應用力學研究所 | zh_TW |
顯示於系所單位: | 應用力學研究所 |
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