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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 劉振軒(Chen-Hsuan Liu) | |
dc.contributor.author | Kuo-Hao Chiu | en |
dc.contributor.author | 邱國皓 | zh_TW |
dc.date.accessioned | 2021-06-15T01:37:13Z | - |
dc.date.available | 2010-07-23 | |
dc.date.copyright | 2009-07-23 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-07-16 | |
dc.identifier.citation | Aly, M. M., Smith, E. J. and Fadly, A. M. (1993). Detection of reticuloendotheliosis virus infection using the polymerase chain reaction. Avian Pathol, 22, 543-554.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/43106 | - |
dc.description.abstract | 禽類淋巴球增生性腫瘤性疾病,特別是淋巴瘤常發生於多種禽類品種,此些病造成雞隻之死亡及產蛋率下降,在全球養雞產業上造成極大的經濟衝擊。大部分的禽類淋巴瘤多與馬立克病病毒(Marek’s disease virus, MDV)家禽白血病病毒(Avian leukosis virus, ALV)及網狀內皮細胞增生症病毒(Reticuloendotheliosis virus, REV)有關。這三個病毒不僅影響禽類產業且同時也造成了多種瀕臨絕種野生鳥類的滅絕,此外這三種疾病沒有典型的病理區別診斷證據可供參考,造成了診斷上極大的困難。研究上通常使用免疫組織化學染色(immunohistochemistry, IHC)、原位雜合技術(in situ hybridization, ISH)和聚合酶鏈鎖反應(polymerase chain reaction, PCR)作為診斷工具。基於上述本研究主要目的為使用PCR及IHC方式進行前述三種病毒引起之禽類淋巴瘤區別診斷。在本研究中,共收集了103個禽類淋巴瘤的組織樣本,其中包括93例石蠟包埋組織塊及10例新鮮病材;包括神經系統、心血管系統、消化道系統、泌尿生殖系統及淋巴系統。PCR方面顯示分別在ALV, MDV及REV檢測出55.43%, 21.36%, 和 21.36%的陽性結果。IHC方面使用病毒抗體分別在ALV, MDV及REV檢測出75.81%, 75.81%, 和 61.29% 的陽性結果。結果顯示IHC在禽類淋巴瘤之石蠟包埋組織塊是一項最有效的檢測工具,但是在發展成熟的淋巴瘤細胞卻檢測不到病毒的抗原蛋白。ISH的建立成功幫助了本研究中在IHC陰性的腫瘤細胞中偵測到REV的病毒核酸。由結果顯示,合併使用IHC及ISH技術在回溯性調查研究中為診斷禽類淋巴球增生性腫瘤性疾病最具有效性的方法。 | zh_TW |
dc.description.abstract | Avian lymphoproliferative neoplastic diseases, especially lymphoma often occurs in a variety of avian species and have economic impact due to associated high mortality and decreased egg production rate. Most of the avian lymphomas are associated with Marek's disease virus (MDV), avian leukosis virus (ALV) or reticuloendotheliosis virus(REV). These three viruses not only influence the poultry industry but also affect a variety of endangered species of wild birds and often lack of classical pathological diagnostic feature. Therefore, immunohistochemistry (IHC), in situ hybridization (ISH), and polymerase chain reaction (PCR) have been developed as diagnostic tools. Based on the purpose aforementioned, the aim of this study is to use PCR and IHC to differentiate the three virus-induced lymphomas. In this study, a total of 103 avian lymphoma tissue samples collected including 93 cases of paraffin-embedded tissue and 10 cases of fresh samples. All samples comprised of the nervous system, cardiovascular system, gastrointestinal system, genito-urinary system and the lymphatic system. The positive results of PCR assay show 55.43%, 21.36%, and 21.36% in ALV, MDV, and REV retrospectively. The IHC using viral antibodies shows 75.81%, 75.81%, and 61.29% in ALV, MDV, and REV retrospectively. In conclusion, IHC employed in formalin-fixed paraffin-embedded tissues of avian lymphoid neoplasms is the most effective diagnostic tool among the three diagnostic tools, especially in retrospective studies; however, viral antigens are less detected among the well-developed tumor cells of ALV or REV induced lymphomas. Establishment of ISH has successfully detected the REV nucleic acid in well-developed tumor cells in this study. By the results showed that the combination of using IHC and ISH has a great potential in retrospective study for the diagnosis of the three viruses associated avian lymphoproliferative neoplasms. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T01:37:13Z (GMT). No. of bitstreams: 1 ntu-98-R96629002-1.pdf: 4898817 bytes, checksum: 94b006326f5c4c8a82e4ac95948aa9b1 (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 中文摘要 I
英文摘要 II 中文摘要 I 英文摘要 II 目錄 III 表目錄 VI 圖目錄 VII 第一章 前言 1 第二章 文獻探討 3 2.1 病毒引起之腫瘤性疾病 3 2.2 馬立克病(Marek’s disease, MD) 9 2.2.1 歷史背景 9 2.2.2 病毒特性 10 2.2.3 臨床特徵及肉眼病變 10 2.2.4 組織病變 11 2.2.5 致癌機轉 12 2.3 家禽白血病(Avian leukosis, AL) 14 2.3.1 歷史背景 14 2.3.2 臨床特徵及肉眼病變 15 2.3.3 組織病變 16 2.3.4 致癌機轉 16 2.4 網狀內皮細胞增生症(Reticuloendotheliosis, RE) 17 2.4.1 歷史背景 17 2.4.2 病毒特性 17 2.4.3 臨床特徵及肉眼病變 18 2.4.4 組織病變 19 2.4.5 致癌機轉 19 2.5 免疫組織化學染色(Immunohistochemistry; IHC) 20 2.5.1 免疫組織化學染色方法原理 20 2.5.1.1 單株及多株抗體之特性 20 2.5.1.2 抗原復性 (antigen retrieval) 21 2.5.1.3 抗原偵測系統 (detection system) 22 2.6 聚合酶鏈鎖反應(Polymerase chain reaction; PCR) 24 2.6.1 聚合酶鏈鎖反應之發展 24 2.6.2 聚合酶鏈鎖反應之原理 25 2.6.3 聚合酶鏈鎖反應之基本要素 26 2.7 原位雜合反應(In Situ hybridization; ISH) 27 2.7.1 原位雜合反應之原理 27 第三章 材料與方法 29 3.1 實驗設計及流程 29 3.2 禽類淋巴增生性腫瘤性疾病病例之收集 30 3.3 外觀及肉眼解剖病變檢查 30 3.4 組織病理學檢查 30 3.5 免疫組織化學染色 32 3.6 分子生物學檢測 36 3.6.1 病毒核酸及前病毒之萃取 36 3.6.2 16s PCR診斷 37 3.6.3 病毒引子對選擇(Table 3.2) 37 3.6.3.1 馬立克病病毒之引子對 37 3.6.3.2 家禽白血病病毒之引子對 37 3.6.3.3 網狀內皮細胞增生症病毒之引子對 38 3.6.4 聚合酶鏈鎖反應(PCR) 38 3.6.4.1 16S PCR 38 3.6.4.2 馬立克病病毒、 白血病病毒及 網狀內皮細胞增生症病毒之PCR檢測 39 3.6.5 設備、材料及藥品試劑 40 3.7 聚合酶鏈鎖反應產物之選殖(cloning) 41 3.7.1 PCR產物之膠體純化 41 3.7.2 使用TA-cloning kit進行病毒核酸之選殖 41 3.7.3 質體萃取 42 3.8 原位雜合反應 43 3.8.1 核探針合成 43 3.8.2 免疫墨點法(Immuno-blot) 43 3.8.3 原位雜合反應步驟 43 第四章 結果 45 4.1 禽類淋巴增生性腫瘤性疾病樣本收集 45 4.2 形態病理學檢查 45 4.2.1 外觀及肉眼病變 45 4.2.2 顯微變化 45 4.3 免疫組織化學染色 48 4.3.1 馬立克病 48 4.3.2 家禽白血病 48 4.3.3 網狀內皮細胞增生症 49 4.4 分子生物學檢查 49 4.5 原位雜合技術建立 49 第五章 討論 68 5.1 以樣本年齡分佈分析 68 5.2 分子生物學檢測討論 69 5.3 組織病理結果與免疫組織化學染色討論 70 5.4 腫瘤細胞形態學區別 74 5.5 總結及未來發展目標 74 第六章 參考文獻 77 | |
dc.language.iso | zh-TW | |
dc.title | 以分子生物學及病理學方法診斷禽類淋巴球增生性腫瘤疾病 | zh_TW |
dc.title | Molecular and Pathological Diagnoses on Avian Lymphoproliferative Neoplasms | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 王金和(Ching-Ho Wang) | |
dc.contributor.oralexamcommittee | 張清棟,李進成 | |
dc.subject.keyword | 禽類,淋巴球增生性腫瘤性疾病,免疫組織化學染色,聚合酶,鏈鎖反應,原位雜合技術, | zh_TW |
dc.subject.keyword | Avian,lymphoproliferative neoplastic diseases,immunohistochemistry,polymerase chain reaction,in-situ hybridization, | en |
dc.relation.page | 97 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2009-07-16 | |
dc.contributor.author-college | 獸醫專業學院 | zh_TW |
dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
顯示於系所單位: | 獸醫學系 |
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