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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳媺玫 | |
| dc.contributor.author | Pei-Shan Hsieh | en |
| dc.contributor.author | 謝佩珊 | zh_TW |
| dc.date.accessioned | 2021-06-17T00:55:14Z | - |
| dc.date.available | 2014-10-21 | |
| dc.date.copyright | 2011-10-21 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-09-26 | |
| dc.identifier.citation | 徐達。(2007) 錦鯉疱疹病毒傳播途徑之探討。國立台灣大學獸醫學研究所碩士論文。.
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(1985) Detection of channel catfish virus in asymptomatic adult channel catfish, Ictalurus punctatus (Rafinesque)*. Journal of Fish Diseases, 8, 485-493. Wittmann G. & Rziha H. (1989) Aujeszky's disease (pseudorabies) in pigs. Developments in veterinary virology (USA). Wolf H., Haus M. & Wilmes E. (1984) Persistence of Epstein-Barr virus in the parotid gland. Journal of virology, 51, 795. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66748 | - |
| dc.description.abstract | 錦鯉疱疹病毒 (koi herpesvirus, KHV)，目前被分類於鯉魚疱疹病毒第三型 (Cyprinid herpesvirus 3, CyHV-3)，為具封套雙股DNA病毒，其核酸大小為295kbp。會造成錦鯉 (Cyprinus carpio koi)與鯉魚 (Cyprinus carpio carpio) 80-100%的死亡率，造成經濟上極大損失；病毒最佳感染溫度為18-28℃。當細胞培養溫度上升至30℃時，病毒會開始停止複製且基因停止轉錄。而在臨床上，當溫度低於18℃或高於28℃時，受感染魚隻臨床症狀會消失並造成潛伏感染；而這些帶原魚可能會在季節變化時，將體內潛伏感染之KHV傳播給健康魚隻，而造成大規模的疾病爆發。而本實驗室以原位雜交 (in situ hybridization) 可在發病魚的巨噬細胞核與質中偵測到病毒核酸；且在2008年翁的實驗中，以KHV病魚乳劑感染健康錦鯉的巨噬細胞，透過Real-time PCR技術，可發現病毒核酸隨時間增加而上升。故初步認為KHV可在巨噬細胞中複製。
因此本實驗主要研究目的為偵測KHV是否會在帶原魚隻血球細胞中潛伏，並利用溫度變化以及ConA刺激，偵測是否表現或合成完整病毒顆粒；根據實驗結果顯示，無論健康魚或帶原魚在23℃下進行攻毒後7-13天，血液中會有病毒早期與晚期的mRNA表現，而在32℃下則僅有早期mRNA表現，顯示在不適當的感染溫度下病毒在周邊血液白血球中無法完成完整的複製；另外可以發現健康魚在23℃第一次接觸病毒時，不會產生中和力價，且魚隻在攻毒後10天開始出現死亡；而32℃攻毒，魚隻不會出現死亡的現象但也沒有中和力價的出現；另外帶原魚在不同溫度下接觸病毒時，會產生中和力價使魚隻可以耐過感染，顯示帶原魚可以有效耐過病毒的第二次感染；另外以TO與KF-1細胞株進行KHV攻毒，可以發現對KHV不具感受性之細胞僅會表現早期的mRNA，而不會出現晚期mRNA之表現。感染KHV後的TKF則會表現早期與晚期mRNA。 | zh_TW |
| dc.description.abstract | Koi herpesvirus (KHV), designated Cyprinid herpesvirus 3 (CyHV-3), is the etiological agent of an emerging and mortal disease in koi (Cyprinus carpio koi) and common carp (Cyprinus carpio carpio). The genome comprises a linear double-stranded DNA of 295 kbp. The disease is seasonal; afflict fish when the water temperature is 18 to 28°C. Virus propagation and virus gene transcription are turned off when cell are moved to a nonpermissive temperature of 30°C. And CyHV-3 can persist asymptomatically for long periods in the fish if the temperature out of optimal temperature or latency sustained, bursts of new infection occur after exposure to permissive temperatures and stress. Our previous data showed that thymidine kinase gene of KHV was detected in macrophages of infected koi by in situ hybridization and Weng found out that KHV TK gene copy number increased in the head kidney leukocyte derived from koi when detected by real-time PCR.
In our study, experimental result showed out, the mRNA of Orf4 (early transcription gene) and Orf2 (late transcription gene) can be detected both in KHV infected naive and carrier carp at 7-13 days postinfection (d.p.i.) when temperature at 23°C. But at 32°C only Orf4 could be detected, demonstrated the virus is unable to complete the replication in peripheral blood leucocytes (PBL) under nonpermissive temperature. In naive carp immersed with KHV the peak of the mortality at 10 d.pi. and serum neutralization (SN) titer can’t be detected within this duration at 23°C. Howerer at 32°C, the naive carp immersed with KHV showed normal appearance without death. The carrier carp exposed to KHV in both permissive and nonpermissive temperature appeared no death and higher SN titer. Moreover only Orf4 could be detected in TO and KF-1 that appeared not susceptibile to KHV, and both Orf4 and Orf2 can be detected in KHV infected TKF. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T00:55:14Z (GMT). No. of bitstreams: 1 ntu-100-R98629029-1.pdf: 1955508 bytes, checksum: 9068ea5da24f83e1121b590a26a74659 (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 致謝 II
中文摘要 III ABSTRACT IV 目錄 V 表目錄 IX 圖目錄 X 第一章 緒論 1 第二章 文獻回顧 3 第一節 錦鯉疱疹病毒之簡介 3 1.1 病毒型態與命名分類 3 1.2 病毒之分子結構與複製 3 1.3 宿主與發病因子 6 1.4 臨床症狀、剖檢病變與病理變化 6 1.5 傳染途徑與傳播方式 7 1.6 診斷方式 8 1.7 病毒的表現複製與新病毒的釋出 12 第二節 魚類的免疫學 14 2.1 魚類免疫主要器官 14 2.2 病毒感染宿主後的免疫反應 14 2.3 非特異性免疫反應 15 2.4 特異性免疫反應 16 2.5 溫度與魚隻免疫之關係 17 第三節 疱疹病毒感染與周邊血液白血球之關係 19 3.1 Alphaherpesvirinae 19 3.2 Betaherpesvirinae 20 3.3 Gammaherpesvirinae 21 3.4 Alloherpesviridae 21 第四節 其他魚類病毒與周邊血液白血球之關係 22 第三章 材料與方法 24 第一節 實驗設計 24 第二節 細胞培養與病毒繼代 25 2.1 實驗材料 25 2.2 病毒增殖與繼代 26 2.3 細胞半數感染量 (50% tissue culture infectious dose, TCID50) 26 2.4 病毒斑試驗 (Plaque assay) 26 2.5 病毒分離 (virus isolation) 27 第三節 病材來源與飼養 27 第四節 攻毒前檢測 27 第五節 魚隻攻毒與腹腔注射DEXAMETHASONE 28 第六節 飼養魚水溫的改變 28 第七節 死亡魚隻的檢測 29 第八節 魚隻的監測 29 8.1 魚隻定期採樣 29 8.2 去氧核醣核酸(DNA)之萃取 30 8.3 聚合酶鏈鎖反應 (polymerase chain reaction, PCR) 31 8.4 巢式聚合酶鏈鎖反應 (Nested polymerase chain reaction, Nested PCR) 31 8.5 瓊脂醣凝膠電泳 (Agarose gel electrophoresis) 32 8.6 病毒中和試驗 (Neutralizing test) 33 8.7 周邊血液白血球的分離 34 8.8 RNA萃取 35 8.9 cDNA的合成 36 8.10 RT-PCR(反轉錄-聚合酶鏈反應) 36 第九節 CON A與LPS刺激對周邊血液細胞中KHV表現的影響 38 第十節 不同細胞對KHV之感受性比較 38 第四章 實驗結果 40 第一節 攻毒前檢測 40 第二節 魚隻死亡率與死亡時間 40 第三節 死亡魚隻病原和抗體檢測 40 第四節 死亡魚隻病毒分離 41 第五節 PCR、NESTED PCR結果 41 第六節 病毒中和試驗 42 第七節 CON A刺激對周邊血液細胞中KHV表現的影響 43 第八節 RT-PCR 43 第九節 不同細胞對KHV之感受性比較 44 9.1 RT-PCR 44 9.2 病毒斑 44 第五章 討論 56 第一節 魚隻死亡率與死亡時間 56 第二節 中和抗體力價之監控 58 第三節 帶原魚體內之病毒再活化 60 第四節 CON A與周邊血液白血球之關係 62 第五節 周邊血液白血球與KHV的關係 63 第六節 不同細胞對KHV之感受性比較 65 第六章 參考文獻 67 | |
| dc.language.iso | zh-TW | |
| dc.title | 探討錦鯉疱疹病毒在潛伏感染錦鯉所分離之周邊血液白血球細胞中之表現 | zh_TW |
| dc.title | Expression of Koi Herpesvirus in Peripheral Blood Leucocyte Cells Isolated from Latently Infected Koi | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 涂堅,謝嘉裕,張本恆 | |
| dc.subject.keyword | 錦鯉疱,疹病毒,RT-PCR,中和抗體,帶原, | zh_TW |
| dc.subject.keyword | KHV,CyHV-3,RT-PCR,serum neutralization titer,carrier, | en |
| dc.relation.page | 77 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-09-28 | |
| 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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