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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46919完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 蕭世烜(Shih-Hsuan Hsiao) | |
| dc.contributor.author | Chi-Chun Tseng | en |
| dc.contributor.author | 曾琪鈞 | zh_TW |
| dc.date.accessioned | 2021-06-15T05:43:19Z | - |
| dc.date.available | 2010-08-20 | |
| dc.date.copyright | 2010-08-20 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-20 | |
| dc.identifier.citation | References
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46919 | - |
| dc.description.abstract | 山羊關節腦炎 (Caprine arthritis encephalitis) 是由一種屬於慢病毒屬 (Lentivirinae genus)，反轉錄病毒科 (Retroviridae family) 的RNA病毒所引起。此疾病在臨床上所造成的病理變化有非化膿性免疫媒介性的關節炎、白質腦炎、間質性肺炎以及乳房炎。為了評估目前此疾病在台灣的感染情況，我們使用商業化的ELISA套組檢測來自羊場的樣本。另外，為了評估抗反轉錄病毒藥物使用於山羊關節腦炎病毒感染控制計畫中的可能性，我們建立了一個可讓病毒存活的細胞外模式，此模式採用血清學陽性羊隻羊乳的體細胞所做的初代培養，另外配合使用可增幅山羊關節腦炎病毒env provirus片段的引子對和探針所建立的可定量即時聚合酶鏈鎖反應以監控病毒量。在2332個山羊血清及乳汁當中，血清學陽性比率為67.30%，其中所有被檢測羊場都是陽性場。乳羊場的血清學陽性比率比肉羊場來的高，而母羊的血清學陽性比率比公羊高，於2007年以後出生的羊隻血清學陽性比率明顯較之前的來的低。來自不同羊隻的山羊乳汁體細胞初代培養，其內的病毒變化曲線之型態不盡相同，所以使用不同來源乳汁體細胞初代培養時實驗內必須加入相同來源的控制組。三株投與stavidine的初代培養中有兩株與未投藥組有顯著差異，而三株投與zidovudine的初代培養中有一組與未投藥組有顯著差異。 | zh_TW |
| dc.description.abstract | Caprine arthritis-encephalitis (CAE) is caused by a RNA virus of Lentivirinae genus, Retroviridae family. The disease is clinically and pathologically manifested by nonsuppurative immune-mediated arthritis, leukoencephalitis, interstitial pneumonia and mastitis. To evaluate CAE infection status in Taiwan, a commercialized ELISA kit was selected to exam goat serum and milk as well as tank milk samples from several selected and voluntary goat herds. In addition, an in vitro model of CAEV propagation was established to investigate the efficacy of anti-retroviral agents, zidovudine and stavudine, on CAE virus propagation in caprine milk primary somatic cell culture. Data indicated that among 2332 serum and milk samples, 67% were serologically positive. 129 tank milk samples were all seropositive, in other words, none of the goat herds were serologically negative. The positive rates were higher for dairy goats, female goats, and goats born after 2008 than others. Additionally, real-time PCR for CAE provirus env gene was able to detect at 101 copy number of the target gene. Both zidovudine and stavudine had no significant cytotoxicity between 1mM to 0.1 μM. Among 3 batches of caprine milk primary somatic cell culture, stavudine treatment significantly suppressed CAE virus propagation in 2 of the batches, and zidovudine suppressed 1 batch. Therefore, anti-retroviral agents might be effective on suppressing CAE virus propagation. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T05:43:19Z (GMT). No. of bitstreams: 1 ntu-99-R97629032-1.pdf: 1387532 bytes, checksum: 66fce6f7f393b5b3e68b3a195bf44c1b (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | Contents
口試委員會審定書…………………………………………………………………Ⅰ 致謝…………………………………………………………………………………Ⅱ 摘要………………………………………………………………………………… Ⅲ Abstract………………………………………………………………………………Ⅳ Contents………………………………………………………………………………Ⅴ Figured………………………………………………………………………….……Ⅷ Tables…………………………………………………………………………………Ⅸ Chapter 1: Literature review……………………………………………………...……1 1.1 Background of caprine arthritis encephalitis virus……………….………1 1.1.1 History………………………………………………………………1 1.1.2 Epidemiology…………………………………………….…………2 1.1.3 Transmission pathway………………………………………………2 1.1.4 Clinical signs………………………………...……………...………4 1.1.5 Pathogen biological characteristic………………………..…………6 1.1.5.1 Virion structure……………………………………………….……6 1.1.5.2 Life cycle………………………………………………………..…8 1.1.5.2.1 Cell and tissue tropism…………………………….……………8 1.1.5.2.2 Cell binding and membrane fusion…………………………..…9 1.1.5.2.3 Nuclear entry of reversetranscripted viral DNA………………10 1.1.5.2.4 Transcription of provirus………………………………………11 1.1.5.2.5 Translation and virion maturation………………..……………12 1.2 Diagnostic assay of CAE virus infection………………………….……13 1.2.1 Serological tests……………………………………………………14 1.2.1.1 ELISA test……………………………………………..…………15 1.2.2 Virus detection tests………………………………..………………15 1.2.2.1 PCR reactions………………………………………………….…16 1.3 Management of CAE virus infection……………………………………17 1.3.1 Total replacement………………………………………….………18 1.3.2 Neonates isolation…………………………………………………19 1.4 Economic consequences of CAE virus infection……………….………20 1.5 CAE virus infection in Taiwan…………………………………….……22 1.6 Antiretroviral compounds………………………………………………23 Chapter 2: Aims and hypothesis……………………………………………...………25 2.1 Difficulties of Taiwan…………….…………………………………..…25 2.1.1 Control programs of CAE virus infection at present are spending long time to achieve virus-free status………………………...……25 2.1.2 High serological prevalence makes CAE virus control program difficult to put into practice……………………………………..…25 2.2 Aims…………………………………………………………….………26 2.2.1 Evaluate the present serological prevalence of CAE virus infection in Taiwan………………………………………………………..…26 2.2.2 Estimate the feasibility of applying antiretroviral compounds in CAE virus infection control program……………………...………26 2.3 Hypothesis………………………………………………………………26 2.3.1 Serological prevalence has maintained or became higher compare with before…………………………………………………………27 2.3.2 Under treatment of antiretroviral compound, CAE virus replication is inhibited…………………………………………………………27 2.4 Rationales…………………………………………………………….…27 2.4.1 Result of ELISA test for the field goat serum or milk samples can reflect the CAE virus infection status in the herds……………...…27 2.4.2 Result of CAE virus env provirus fragment amplified by quantitative Real-time PCR can represent the viral load of CAE virus……...…28 Chapter 3: Materials and methods……………………………………………………29 3.1 Serological surveillance…………………………………………….....…29 3.1.1 Sample collection and preparation…………………………...….…29 3.1.2 CAE ELISA test………………………………………………...…29 3.2 Antiretroviral compounds selecting in vitro model establishment…..….30 3.2.1 Real-time polymerase chain reaction……………………...………30 3.2.1.1 Positive control establishment……………………………………30 3.2.1.1.1 Sample selection……………………………………….………30 3.2.1.1.2 Nucleic acid extraction……………………………...…………30 3.2.1.1.3 Real-time PCR reaction procedure……………………….……32 3.2.1.1.4 TA cloning…………………………………...…………...……32 3.2.2 In vitro model establishment………………………………………32 3.2.2.1 Goat milk somatic and collection……………………...…………32 3.2.2.2 Primary culture procedure………………………………..………33 3.2.2.3 Immunofluorescence assay……………………………….………33 3.2.3 Observation of in vitro model……………………………..………34 3.2.3.1 Monitoring of viral load………………………….…..…..………34 3.2.3.2 Monitoring of cell number……………………………….………34 3.2.4 Antiretroviral compounds screening………………………………35 3.2.4.1 Antiretroviral compounds preparations………………..…………35 3.2.4.2 Cytotoxicity evaluation………………………………………..…35 3.2.4.3 Antiretroviral efficacy evaluation…………………….…….……36 Chapter 4: Results……………………………………………………………………38 4.1 Serological surveillance…………………………………………...……38 4.1.1 No seropositive goat herd was found among tested herds. Lower seropositive percentage for breeding goat herds than others.……..……..……..……..……..……..……..……..……..…38 4.1.2 Seropositve percentage results from different gender, age, county and breed………..……..……..……..…………………………..…38 4.2 Established Real-time PCR is able to detect the virus from 10 to108 copy number…………………………………………………………….……39 4.3 Primary culture of goat milk somatic cell………………………………39 4.4 Positive signal of CAE virus capsid protein and cytokeratin-18 are detected in Immuno fluorescence assay (IFA) …………………………39 4.5 Zidovudine and stavudine have no significant cytotoxicity effect to goat milk somatic primary culture……………………………………...……40 4.6 Viral load variation pattern are inconsistent for different origin goat milk somatic cells primary culture………………………………...…………40 4.7 There are significant differences of viral load between Mock, Zidovudine and Stavudine treated groups. ……………………………...…………40 Chapter 5: Discussion……………………………………………………………..…53 Chapter 6: Conclusion…………………………………………………….…….……60 References……………………………………………………………………………61 Addendum | |
| dc.language.iso | en | |
| dc.subject | 抗反轉錄病毒藥物 | zh_TW |
| dc.subject | 山羊關節腦炎病毒 | zh_TW |
| dc.subject | 血清學調查 | zh_TW |
| dc.subject | 台灣 | zh_TW |
| dc.subject | 山羊乳汁體細胞初代培養 | zh_TW |
| dc.subject | Taiwan | en |
| dc.subject | antiretroviral compound | en |
| dc.subject | goat milk somatic cells primary culture | en |
| dc.subject | CAE virus | en |
| dc.subject | serological surveillance | en |
| dc.title | 台灣山羊關節腦炎血清學檢測與在初代培養山羊乳汁體細胞中探討抗反轉錄病毒藥物對抗山羊關節腦炎病毒增殖之效力 | zh_TW |
| dc.title | Serological Survey of Caprine Arthritis Encephalitis (CAE) in Taiwan and Efficacy of Anti-Retroviral Agents on CAE Virus Propagation in Caprine Milk Primary Somatic Cell Culture | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 龐飛(Victor Fei Pang),鄭謙仁(Chian-Ren Jeng) | |
| dc.contributor.oralexamcommittee | 劉涓,張志誠 | |
| dc.subject.keyword | 山羊關節腦炎病毒,血清學調查,台灣,山羊乳汁體細胞初代培養,抗反轉錄病毒藥物, | zh_TW |
| dc.subject.keyword | CAE virus,serological surveillance,Taiwan, goat milk somatic cells primary culture,antiretroviral compound, | en |
| dc.relation.page | 92 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-08-20 | |
| 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 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-99-1.pdf 未授權公開取用 | 1.36 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。