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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76681完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 何佳安(Ja-an Annie Ho) | |
| dc.contributor.author | You-Jin Chen | en |
| dc.contributor.author | 陳幼瑾 | zh_TW |
| dc.date.accessioned | 2021-07-10T21:34:58Z | - |
| dc.date.available | 2021-07-10T21:34:58Z | - |
| dc.date.copyright | 2016-11-02 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-08-17 | |
| dc.identifier.citation | 1. Kueppers, R., The biology of Hodgkin's lymphoma. Nat. Rev. Cancer. 2009, 9 (1), 15-27.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76681 | - |
| dc.description.abstract | 微型核糖核酸(MicorRNA, miRNA)為長約22個核苷酸片段，高度保留，非編碼的RNA分子；可藉由配對信使RNA (Messenger RNA)的三端不轉譯區域調控基因表現。MicroRNAs在生物體內扮演重要角色，影響細胞的增生、分化、凋亡及代謝。許多證據顯示，不正常表現的microRNA可藉由調節腫瘤抑癌或致癌基因影響癌症進程。近來研究發現，哺乳類惡性淋巴瘤中，有一種常見的侵襲性非何杰金氏B細胞淋巴瘤(Non-Hodgkin’s B-cell lymphoma)之亞型─瀰漫性大型B細胞淋巴瘤(Diffuse large B-cell lymphoma)，有miR-155高度表現的證據。因此，miR-155被認為是有潛力做為淋巴瘤的生物標記核酸分子。本研究中，我們提出一個新的檢測策略，結合Duplex-specific nuclease (DSN)輔助目標分子循環再生(Target recycling)與鏈置換擴增(Strand displacement amplification, SDA)技術，並以電化學分析法偵測樣品中miR-155含量。我們首先以聚丙烯醯胺膠體電泳法證明經由DSN作用後所獲得的核酸片段，足可啟動SDA，並產生訊號誘導序列。接著確定此誘導序列可與電極表面上亞甲藍標定的髮夾型探針進行雜合，進而產生電化學訊號之差異性。以“訊號關閉”的電化學生物感測器，提供檢測miR-155的可行性與潛力，可做為惡性淋巴瘤之液態樣品切片(Liquid biopsy)的臨床診斷工具。 | zh_TW |
| dc.description.abstract | MicroRNAs (miRNAs) are a class of small (~22nt), highly conserved, non-coding RNA that negatively regulate gene expression through pairing the 3’ untranslated region of mRNA transcripts. MicroRNAs play a critical role in several cellular regulation such as proliferation, differentiation, apoptosis, and metabolism. Growing evidences have shown that aberrant expression of miRNAs links to various cancers by modulating the expression of tumor suppressors or oncogenes. Recent studies showed that microRNA-155 (miR-155) was up-regulated in mammalian B-cell malignancies, such as diffuse large B cell lymphoma (DLBCL), which is an aggressive type of non-Hodgkin’s lymphoma. MiR-155 is hence suggested as a potential biomarker for lymphoma. Herein, we proposed a new strategy for electrochemical detection of miR-155 based on duplex-specific nuclease (DSN)-assisted target recycling and strand displacement amplification (SDA). Our preliminary results collected by polyacrylamide gel electrophoresis (PAGE) confirmed the production of trigger sequences through DSN reaction, followed by the onset of the SDA, leading to the production of the signal inducer sequences. Finally, signal inducer sequences hybridized with the hairpin probes on electrode surface sensitized with methylene blue, resulting in the establishment of a ‘signal-off’ electrochemical biosensor. This biosensing platform provides a promising applicability and feasible potential toward the detection of miR-155 that can be further extended to clinical diagnosis of liquid biopsy samples for patients with mammalian lymphoma. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-10T21:34:58Z (GMT). No. of bitstreams: 1 ntu-105-R03B22031-1.pdf: 3394352 bytes, checksum: 2977d9578a7e532616e7a35ad8f8f5e1 (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 第一章 緒論 1
1.1. 淋巴瘤 (Lymphoma) 1 1.1.1 人類淋巴瘤簡介 1 1.1.2 犬淋巴瘤簡介 1 1.1.3 淋巴瘤診斷 5 1.2. 微型核糖核酸 (MicroRNA) 6 1.2.1 microRNA生合成與作用機制 6 1.2.2 microRNA與癌症 8 1.2.3 miR-155作為淋巴癌的生物指標 10 1.2.4 microRNA的檢測方法 10 1.3. 酵素輔助核酸目標循環作用 12 1.3.1 Duplex-specific nuclease輔助目標物循環法 13 1.4. 等溫核酸放大技術 14 1.4.1 單鏈置換擴增法 (Strand displacement amplification, SDA) 14 1.5. 電化學DNA感測器 16 1.6. 電化學偵測法 18 1.7. 研究動機 21 第二章 實驗材料與方法 22 2.1. 核酸序列 22 2.2. 實驗試劑與材料 23 2.3. 實驗儀器 26 2.4. 緩衝溶液 27 2.5. 聚丙醯胺膠體電泳法 (Polyacrylamide gel electrophoresis, PAGE) 28 2.6. DSN輔助目標物循環與單鏈置換擴增法之反應 28 2.6.1. DSN酶切產物探討 28 2.6.2. DSN輔助目標物循環法之測試 28 2.6.3. DSN輔助目標物循環法之最佳化 29 2.6.4. DSN高溫去活反應 29 2.6.5. DSN之目標分子專一性酶切測試 29 2.6.6. 單鏈置換擴增法之測試 30 2.6.7. 單鏈置換擴增法之最佳化 30 2.7. 鍍金時間對MB-hairpin DNA電流訊號之影響 31 2.8. 電解質緩衝液選擇 33 2.9. MB-hairpin DNA修飾於AuNP-SPCE之鑑定 34 2.10. SPCE、AuNP-SPCE與DNA@AuNP-SPCE之電極表面鑑定 36 2.11. 全流程反應 37 第三章 實驗結果與討論 38 3.1 實驗設計 38 3.2 DSN輔助目標分子之循環反應(DSN-assisted target recycling) 41 3.2.1. DSN酶切產物之探討 41 3.2.2. DSN可輔助目標分子進行循環反應 44 3.2.3. DSN反應溫度最佳化 45 3.2.4. DSN使用濃度最佳化 47 3.2.5. DSN加熱處理去活化時間之最佳化 49 3.2.6. DSN之專一性酶切目標分子測試 51 3.3 單鏈置換擴增法(Strand displacement amplification, SDA) 53 3.3.1. 單鏈置換擴增法測試 53 3.3.2. 反應使用酵素比例最佳化 55 3.4 工作電極製備及其最佳化 57 3.4.1. 鍍金時間對MB-hairpin DNA電流訊號之影響 57 3.4.2. 電解質緩衝液選擇 59 3.4.3. MB-hairpin DNA修飾於AuNP-SPCE之鑑定 61 3.4.4. SPCE、AuNP-SPCE、DNA@AuNP-SPCE之電極表面鑑定 63 3.5 全流程反應 66 第四章 結論 70 第五章 參考文獻 72 | |
| dc.language.iso | zh-TW | |
| dc.subject | 恆溫放大法 | zh_TW |
| dc.subject | 淋巴瘤 | zh_TW |
| dc.subject | 液態切片樣品 | zh_TW |
| dc.subject | 微型核糖核酸 | zh_TW |
| dc.subject | 電化學感測器 | zh_TW |
| dc.subject | Duplex-specific nuclease | zh_TW |
| dc.subject | electrochemical biosensor | en |
| dc.subject | strand displacement amplification | en |
| dc.subject | duplex-specific nuclease | en |
| dc.subject | lymphoma | en |
| dc.subject | liquid biopsy | en |
| dc.subject | microRNA | en |
| dc.title | 發展觸發式雙重訊號放大策略以偵測與癌症有關之微小核酸核醣片段 | zh_TW |
| dc.title | A Sensitive Electrochemical Detection of a Tumor-associated MicroRNA by Target-triggered Dual Amplification Strategy | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 邢怡銘(I-Ming Hsing),吳立真(Li-Chen Wu),陳平,鄭建中 | |
| dc.subject.keyword | 淋巴瘤,液態切片樣品,微型核糖核酸,電化學感測器,Duplex-specific nuclease,恆溫放大法, | zh_TW |
| dc.subject.keyword | lymphoma,liquid biopsy,microRNA,electrochemical biosensor,duplex-specific nuclease,strand displacement amplification, | en |
| dc.relation.page | 87 | |
| dc.identifier.doi | 10.6342/NTU201602769 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2016-08-18 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科技學系 | zh_TW |
| 顯示於系所單位： | 生化科技學系 | |
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