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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78740完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 沈弘俊(Horn-Jiunn Sheen) | |
| dc.contributor.author | Yung-Yu Huang | en |
| dc.contributor.author | 黃詠榆 | zh_TW |
| dc.date.accessioned | 2021-07-11T15:15:59Z | - |
| dc.date.available | 2025-09-01 | |
| dc.date.copyright | 2020-10-20 | |
| dc.date.issued | 2020 | |
| dc.date.submitted | 2020-08-13 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78740 | - |
| dc.description.abstract | EB病毒是一種與鼻咽癌及淋巴癌高度相關的病毒，而 LMP1是EB病毒中常見的表現蛋白之一，因此本研究便以LMP1 DNA作為檢測標的，利用微流道聚合酶連鎖反應裝置結合奈米狹縫表面電漿共振感測器，開發出一套結合DNA複製及檢測的裝置化整合平台，能夠針對LMP1 DNA及其真實檢體進行免標定、即時、高靈敏度的檢測。 本研究微流道PCR採用分段加溫的方式，可以省去傳統PCR反覆升降溫的多餘能源及時間消耗。感測器使用奈米狹縫SPR晶片，是一種免標定、即時檢測、高靈敏度的生物感測器，由於其體積小、光路設計簡單，因此相當適合與微流道PCR整合。微流道製作方式採用CNC雕刻機雕刻壓克力搭配有機溶液黏合法，成本低且製造快速。奈米狹縫SPR晶片採用自動化奈米壓印技術，於PC板上壓印奈米級光柵結構，製作方式簡單，適合大量製造。 微流道PCR裝置在流速5 g/mL之下，能達到儀器PCR 30 cycles的複製效果，且反應時間僅需要36分鐘，比傳統儀器節省了將近70分鐘。針對低濃度LMP1 DNA的檢測，最低檢測極限濃度達到10 pg/mL，比傳統凝膠電泳檢測方式擁有更低的檢測範圍。真實檢體檢測方面，亦能成功從癌細胞萃取出的DNA中檢測出LMP1基因，顯示本研究裝置應用於真實檢體檢測的可行性。 | zh_TW |
| dc.description.abstract | EB virus is highly associated with Nasopharyngeal carcinoma and Lymphoma, and LMP1 is one of the main performance proteins of the EB virus. A combined device of microfluidic Polymerase Chain Reaction with nanoslit Surface Plasmon Resonance sensor for LMP1 DNA detection has been developed, which enables amplification and detection of LMP1 gene on a combined platform. The combined device performed a label-free, real-time, highly sensitive, low cost, and rapid detection versus LMP1 DNA and its relative real samples. The continued-flow PCR has been used, which can avoid unnecessary energy loss and time consumption, and nanoslit SPR chips have been used as a label-free, real-time, high sensitivity sensor. Considering the small volume and simple optical setup, the nanoslit SPR chip is quite feasible to integrate with microfluidic PCR. Microchannel was fabricated by CNC engraving and solvent bonding of PMMA. Furthermore, automatic Nanoimprinting Lithography has been used to fabricate SPR chips by printing nanoscale grating structure, owning the advantage of low cost and mass production. An optimized flow rate has been found out. For the 30 cycles reaction of PCR, the new method only took 36 minutes, almost 70 minutes less than the conventional method with the same DNA amplification effect. Besides, the different initial concentration of LMP1 DNA has been detected by the combined device. The detection limit of LMP1 DNA is 10 pg/mL. Moreover, the combined device is capable of detecting real samples of extracted DNA from EBV-positive cells, showing the potential of clinical diagnosis. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T15:15:59Z (GMT). No. of bitstreams: 1 U0001-2807202014534500.pdf: 4256285 bytes, checksum: 03f21507f109118a62e27c19ceb352dd (MD5) Previous issue date: 2020 | en |
| dc.description.tableofcontents | 致謝 II 摘要 III Abstract IV 目次 V 圖目次 VII 表目次 10 第一章 導論 11 1.1 前言 11 1.2 研究動機與目的 13 1.3 研究方法 14 1.4 論文架構 15 第二章 文獻回顧 16 2.1 表面電漿共振技術之發展背景 16 2.2 微流道聚合酶連鎖反應之發展背景 19 第三章 實驗原理 23 3.1 表面電漿子原理 23 3.2 表面電漿共振之激發與免疫分析原理 29 3.3 聚合酶連鎖反應原理 32 第四章 實驗架設與方法 34 4.1 PCR微流道與SPR晶片製程 34 4.1.1 PCR微流道製程 34 4.1.2 SPR晶片製程 36 4.2 溫度控制系統 38 4.3 光學偵測系統 40 4.4 PCR配方與引子設計 41 4.5 DNA之萃取 42 4.6 SPR晶片之表面修飾與Probe設計 43 4.7 動態分析方法 45 第五章 實驗結果與討論 46 5.1 微流道尺寸量測 46 5.2 溫控系統之均勻性與穩定性 47 5.3 光學偵測系統搭配SPR晶片之靈敏度量測 48 5.4 整合裝置檢測LMP1 DNA與流速最佳化 49 5.4.1 不同cycle數之LMP1 DNA紅移量 50 5.4.2 不同流速之凝膠電泳量測結果 53 5.4.3 不同流速之SPR量測結果 55 5.4.4 與先前研究之比較 56 5.5 LMP1 DNA之檢測極限 58 5.6 真實檢體之量測 59 第六章 結論與未來展望 64 參考文獻 65 附錄 69 | |
| dc.language.iso | zh-TW | |
| dc.subject | 微流道聚合酶連鎖反應 | zh_TW |
| dc.subject | LMP1 DNA | zh_TW |
| dc.subject | 奈米狹縫表面電漿共振晶片 | zh_TW |
| dc.subject | 免標定即時檢測 | zh_TW |
| dc.subject | LMP1 DNA | en |
| dc.subject | nanoslit SPR | en |
| dc.subject | Microfluidic PCR | en |
| dc.subject | Label-free detection | en |
| dc.title | 微流道聚合酶連鎖反應裝置結合奈米狹縫表面電漿共振感測器用於LMP1 DNA 之檢測 | zh_TW |
| dc.title | Microfluidic Polymerase chain reaction device combined with nanoslit surface plasmon resonance sensor for LMP1 DNA detection | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 109-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 魏培坤(Pei-Kuen Wei) | |
| dc.contributor.oralexamcommittee | 盧彥文(Yen-Wen Lu),范育睿(Yu-Jui Fan) | |
| dc.subject.keyword | LMP1 DNA,免標定即時檢測,微流道聚合酶連鎖反應,奈米狹縫表面電漿共振晶片, | zh_TW |
| dc.subject.keyword | LMP1 DNA,Label-free detection,Microfluidic PCR,nanoslit SPR, | en |
| dc.relation.page | 72 | |
| dc.identifier.doi | 10.6342/NTU202001967 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2020-08-14 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 應用力學研究所 | zh_TW |
| dc.date.embargo-lift | 2025-09-01 | - |
| 顯示於系所單位： | 應用力學研究所 | |
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