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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 萬灼華(Cho-Hua Wan) | |
| dc.contributor.author | Yong-Sin Wei | en |
| dc.contributor.author | 魏詠欣 | zh_TW |
| dc.date.accessioned | 2022-11-23T09:00:12Z | - |
| dc.date.available | 2021-11-05 | |
| dc.date.available | 2022-11-23T09:00:12Z | - |
| dc.date.copyright | 2021-11-05 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-20 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79424 | - |
| dc.description.abstract | "小鼠肝炎病毒（Mouse hepatitis virus, MHV）、小鼠諾羅病毒（Murine norovirus, MNV）及小鼠星狀病毒（Murine astrovirus, MuAstV）是具高傳染力的RNA病毒，常見於小鼠族群。MHV與MNV可能會影響實驗小鼠健康，MuAstV則為新興病毒，此三種病毒皆有潛力影響動物實驗的研究結果，應從實驗小鼠排除，因此有效的診斷方法對於實驗動物管理極為重要。由於飼養管理的改進與IVC系統的使用，使得近年較難穩定地透過衛兵鼠與傳統診斷方法檢測到小鼠病原感染，因此引起後續傳統監測方法的改進與新方法的建立。本研究開發可同時檢測MHV、MNV及MuAstV三種RNA病毒與小鼠管家基因（housekeeping gene）的多重引子聚合酶鏈鎖反應診斷法（MHV/MNV/MuAstV/Actin multiplex RT-PCR assay）。此診斷仍能專一地複製出目標病毒基因，且無論在單獨感染或多重等量感染，最低皆可測得10 copies。模擬三種病毒不等量同時感染的情況下，即使有一種或兩種的高量病毒（104 copies），此試劑仍可檢測到10 ~ 103 copies的其餘病毒。將multiplex RT-PCR用於診斷來自11個台灣研究單位的112個小鼠糞便樣本，顯示本試劑病毒檢測的敏感度為86.2%~100%，準確度為96.4%~100%。此流行率調查結果顯示，MNV陽性率為25.8%，MuAstV的感染率則為51.8%，且無發現MHV感染小鼠。本研究開發之MHV/MNV/MuAstV/Actin multiplex RT-PCR具高專一性與敏感性，能同時診斷多種病原以節省人力、時間及經費，可作為研究單位小鼠定期健康監測之有效診斷方法，且本次病毒流行率調查結果可作為未來疾病控制之參考。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-23T09:00:12Z (GMT). No. of bitstreams: 1 U0001-1810202114121400.pdf: 2604173 bytes, checksum: aa9cf35176d04a54f81880158aead76b (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "口試委員審定書 I 致謝 II 中文摘要 III Abstract IV 目錄 V LIST OF TABLES VII LIST OF FIGURES VIII 第一章、前言 1 1.1研究背景 1 1.2 Mouse hepatitis virus（MHV） 1 1.3 Murine Norovirus（MNV） 3 1.4 Murine Astrovirus（MuAstV） 4 1.5疾病診斷 5 1.6診斷方法 6 1.7血清學（serology）診斷法 7 1.8 分子生物學（molecular biology）診斷法 8 1.9 研究目的 11 第二章、材料與方法（Materials and methods） 13 2.1動物及糞便檢體來源 13 2.2糞便之RNA萃取 13 2.3 核酸引子（Oligonucleotide primer） 14 2.4 RT-PCR（reverse transcription-polymerase chain reaction） 15 2.4.1反轉錄（reverse transcription, RT）反應 15 2.4.2 PCR（polymerase chain reaction） 15 2.5陽性控制組之建立 16 2.5.1 MHV陽性質體 16 2.5.2 MNV陽性質體 18 2.5.3 MuAstV陽性質體 19 2.6 以單一PCR增幅DNA 19 2.6.1 單一病原specific PCR 19 2.6.2單一病原specific PCR之特異性試驗 19 2.6.3單一病原specific PCR之敏感性試驗 20 2.7 MHV/MNV/MuAstV/actin multiplex RT-PCR 20 2.7.1 MHV/MNV/MuAstV/actin multiplex RT-PCR之特異性試驗 20 2.7.2 MHV/MNV/MuAstV/actin multiplex RT-PCR對單一感染之敏感性試驗 20 2.7.3 MHV/MNV/MuAstV/actin multiplex RT-PCR對多重等量感染之敏感性試驗 21 2.7.4 MHV/MNV/MuAstV/actin multiplex RT-PCR對不等量的多病原共同感染之偵測敏感度試驗 21 2.8 MHV、MNA及MuAstV於台灣小鼠設施之流行率調查 22 2.9 DNA定序 22 第三章、結果 23 3.1核酸引子之評估 23 3.2 MHV、MNV或 MuAstV單一病原RT-PCR之專一性 23 3.3 MHV, MNV, MuAstV單一病原RT-PCR之敏感性 24 3.4 MHV/MNV/MuAstV/Actin Multiplex RT-PCR Assay之專一性 24 3.5 MHV/MNV/MuAstV/Actin Multiplex RT-PCR Assay之敏感性 25 3.5.1 Multiplex RT-PCR Assay檢測單獨病原感染之敏感性 25 3.5.2 Multiplex RT-PCR Assay檢測多重病原等量共同感染之敏感性 25 3.5.3 Multiplex RT-PCR Assay檢測多重病原不等量共同感染之敏感性 26 3.6 Multiplex RT-PCR Assay應用於實驗小鼠之調查結果 27 第四章、討論 28 參考文獻 51" | |
| dc.language.iso | zh-TW | |
| dc.title | 小鼠肝炎病毒、小鼠諾羅病毒及小鼠星狀病毒之分子生物診斷試劑之建立 | zh_TW |
| dc.title | "Establishment of a multiplex RT-PCR for detecting MHV, MNV and MuAstV in mice " | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王明升(Hsin-Tsai Liu),廖欽峯(Chih-Yang Tseng) | |
| dc.subject.keyword | 小鼠肝炎病毒,小鼠諾羅病毒,小鼠星狀病毒,多重引子聚合酶鏈鎖反應診斷法, | zh_TW |
| dc.subject.keyword | Mouse hepatitis virus,Murine norovirus,Murine astrovirus,multiplex RT-PCR assay, | en |
| dc.relation.page | 56 | |
| dc.identifier.doi | 10.6342/NTU202103824 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2021-10-21 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 分子暨比較病理生物學研究所 | zh_TW |
| 顯示於系所單位： | 分子暨比較病理生物學研究所 | |
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