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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林啟萬(Chii-Wann Lin) | |
| dc.contributor.author | Wei-Yin Lin | en |
| dc.contributor.author | 林瑋瑩 | zh_TW |
| dc.date.accessioned | 2022-11-25T06:32:54Z | - |
| dc.date.copyright | 2021-11-11 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-25 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82152 | - |
| dc.description.abstract | "自從2019年12月開始，新型冠狀病毒(Severe Acute Respiratory Syndrome Coronavirus 2, SARS-CoV-2)在全球廣泛的傳播，導致確診人數及死亡人數日益攀升。若能夠及早診斷出感染者並進行隔離治療，對於疫情的控制及醫療資源的有效分配將會是一大助益。現行的檢測中，核酸檢測是高準確率且早日有效檢測出病毒的最佳選擇。本研究旨在使用恆溫無酵素放大方法進行SARS-CoV-2基因篩檢的研發，以改善即時逆轉錄聚合酶連鎖反應(real-time reverse transcription-PCR, rRT-PCR)之耗時、需調控溫度、需要酵素等缺點。 恆溫無酵素放大技術在目標序列存在時，會與hairpins結合並產生一連串擴增的現象。本研究使用美國疾管署提出之SARS-CoV-2 N基因探針位點的互補股作為目標序列，進行兩種恆溫無酵素放大技術的序列設計，並使用電泳驗證設計的效果。此外，為了進一步獲得量化的數據，採用高靈敏度、無需標記且即時監測的表面電漿子共振(surface plasmon resonance, SPR)感測器進行優化反應的參數測試、探針對目標序列的特異性測試及臨床應用可行性測試。由結果顯示，優化後的放大訊號可增加50%以上，且加入隨機序列的控制組及加入BSA干擾後，探針與目標序列仍具有高度特異性，驗證具有應用在複雜樣本的潛力。本研究建立了恆溫無酵素放大方法的設計流程、反應優化參數及模擬臨床樣品反應的資訊，提供未來使用此方法相關參照依據。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T06:32:54Z (GMT). No. of bitstreams: 1 U0001-2308202117550400.pdf: 4709201 bytes, checksum: e1f7dc3d5a0d69c351c7317e64866e70 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 誌謝 I 中文摘要 II Abstract III 圖目錄 VIII 表目錄 X 第 一 章 緒論 1 1.1研究背景與重要性 1 1.2研究動機與目的 5 1.3章節架構 6 第 二 章 基本原理與文獻回顧 7 2.1新型冠狀病毒 7 2.1.1新型冠狀病毒介紹 7 2.1.2新型冠狀病毒的數據共享 10 2.1.3現行的標準核酸篩檢方式 11 2.2恆溫無酵素的放大技術 14 2.2.1雜交連鎖反應 14 2.2.2催化髮夾組裝 15 2.2.3恆溫無酵素技術的序列設計 18 2.2.4檢測上的應用 19 2.3表面電漿子共振感測技術 20 2.3.1表面電漿子共振原理 21 2.3.2表面電漿子共振生物感測器 22 2.3.3表面化學修飾方式 23 2.4雜交反應的障礙與排除 25 第 三 章 研究材料與方法 28 3.1反應序列設計及驗證方法 28 3.1.1設計SARS-CoV-2病毒檢測序列 28 3.1.2電泳驗證 28 3.1.3探針序列設計 29 3.2表面電漿子共振系統 29 3.2.1感測晶片的構造 30 3.2.2強度式表面電漿子共振感測器系統架構 30 3.2.3操作介面 32 3.2.3訊號量測與分析 34 3.2.4感測晶片再生方式 35 第 四 章 結果與討論 36 4.1序列資訊 36 4.2電泳印證 38 4.2.1 HCR 38 4.2.2 CHA 40 4.3 優化恆溫無酵素放大反應之SPR量化分析 42 4.3.1探針修飾量與放大反應的量化分析 42 4.3.2 PEG濃度對於訊號的影響 45 4.3.3 stem長度對於訊號的影響 47 4.3.4鹽度對於SPR訊號的影響 48 4.4 探針與目標序列之特異性測試 50 4.5 臨床測試的可行性 50 第 五 章 結論與未來展望 52 第 六 章 參考文獻 54 | |
| dc.language.iso | zh-TW | |
| dc.subject | 表面電漿子共振 | zh_TW |
| dc.subject | 新型冠狀病毒 | zh_TW |
| dc.subject | 恆溫無酵素放大技術 | zh_TW |
| dc.subject | Surface Plasmon Resonance | en |
| dc.subject | Severe Acute Respiratory Syndrome Coronavirus 2 | en |
| dc.subject | Isothermal Enzyme-free Amplification | en |
| dc.title | 恆溫無酵素放大技術於SARS-CoV-2的應用 | zh_TW |
| dc.title | Isothermal Enzyme-free Amplification Targeting on SARS-CoV-2 | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 彭盛裕(Hsin-Tsai Liu),施博仁(Chih-Yang Tseng) | |
| dc.subject.keyword | 新型冠狀病毒,恆溫無酵素放大技術,表面電漿子共振, | zh_TW |
| dc.subject.keyword | Severe Acute Respiratory Syndrome Coronavirus 2,Isothermal Enzyme-free Amplification,Surface Plasmon Resonance, | en |
| dc.relation.page | 63 | |
| dc.identifier.doi | 10.6342/NTU202102641 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2021-08-26 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| dc.date.embargo-lift | 2026-08-25 | - |
| 顯示於系所單位： | 醫學工程學研究所 | |
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