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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳媺玫(Meei-Mei Chen) | |
| dc.contributor.author | Yu-Chiao Yang | en |
| dc.contributor.author | 楊雨樵 | zh_TW |
| dc.date.accessioned | 2021-05-15T17:51:40Z | - |
| dc.date.available | 2017-08-21 | |
| dc.date.available | 2021-05-15T17:51:40Z | - |
| dc.date.copyright | 2014-08-21 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-15 | |
| dc.identifier.citation | 涂堅(2003)。錦鯉疱疹病毒症之防治。行政院農業委員會家畜衛生試驗所。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/5081 | - |
| dc.description.abstract | 錦鯉疱疹病毒 (Koi herpesvirus,KHV),亦稱為鯉魚疱疹病毒第三型 (Cyprinid herpes virus-3, CyHV-3),或鯉魚間質性腎炎及鰓壞死病毒 (carp interstitial nephritis and gill necrosis virus,CNGV),為一具封套的二十面體病毒,歸屬於 Alloherpesviridae科。KHV自1998年於以色列爆發以來,2000年便已在世界各地接連傳出災情,且由世界動物衛生組織(World Organization for Animal Health,OIE)列為水生動物輸出入檢疫疾病。我國亦於2002年12月出現首項確診病例。目前台灣的錦鯉養殖場在季節交替之際,時常傳出KHV感染症的病情報告,但截至目前為止,除了隔離病魚、撲殺與清池外,尚無有效防治手段。因此KHV之疫苗開發刻不容緩。
2003年Ronen等人藉由KHV的連續繼代方式製造減毒疫苗,但由於臨床上無法免除病毒毒力回復之風險,因此限於以色列地區使用。本實驗著眼於死毒疫苗的製作,透過福馬林去活化疫苗的接種,引發錦鯉魚隻的體液免疫反應,並配合弗氏完全佐劑(Freund’s complete adjuvant,FCA)與Montanide ISA 763A兩種佐劑,以增強魚隻的細胞免疫反應,旨在建立魚隻抵抗KHV感染的能力。結果顯示,在Naive魚隻上,未配合佐劑的死毒疫苗可引起高達1:128的血清中和抗體力價,相對存活率(relative percent survival,RPS)亦超過70%;而在Carrier上的RPS甚至可達100%。配合佐劑的疫苗,其血清表現中和抗體力價的時間,較未配合佐劑的超出一倍,在免疫後96天仍可偵測到1:32以上的中和抗體力價。 本實驗成功地在實驗室環境下,透過活體in vivo試驗,確認了KHV福馬林去活化疫苗的效果,未來將進一步將本死毒疫苗之製造條件予以最佳化,以求得更低的成本與更高的保護效率。 | zh_TW |
| dc.description.abstract | Koi Herpesvirus (KHV) which is also called Cyprinid herpes virus-3, (CyHV-3) or carp interstitial nephritis and gill necrosis virus (CNGV) is a icosahedral virus and grouped family Alloherpesviridae. The disease firstly reported in Israel in 1998, then outbreaked around the world until 2000. The huge economical losing has lead KHV to be recognized as certificated disease by World Organization for Animal Health (OIE). In Taiwan, the first case report has been found in December in 2002. The infection of KHV usually took place during season changing in Koi aquaculture field, and there are still no effective control strategies for KHV infection except quarenting, disinfection and pond liming. Therefore the establishment of vaccine is an urgent task to pursue.
Ronen et. al. have developed an attenuated KHV strain as a vaccine through series passaging methods. But the using of attenuated vaccine is totally restricted in Israel area due to the significant virulence recovery risks. This research was mentioned about the Formalin-inactivated method to produce high effective vaccine for inducing humeral immunity of fishes. On the other hand, we also select two adjuvants including Freund’s complete adjuvant (FCA) and Montanide ISA 763A to build up the cell-mediated immunity while vaccing inoculating. After immunizing naive fishes, the high titer serum neutralizing antibodies can be detected, and the relative percent survivol (RPS) was over 70%. And the RPS on Carriers was even approaching 100%. The neutralizing antibodies lasting of inactivated vaccine with adjuvant are two times longer than the vaccine without adjuvant. The titer of serum antibody still reached 1:32 at 96th day post-immunization. We successfully established an effective inactivated KHV vaccine in vivo. We will optimize the producing contidion in order to reduce the cost and raise the efficacy. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-15T17:51:40Z (GMT). No. of bitstreams: 1 ntu-103-R94629014-1.pdf: 6043772 bytes, checksum: e6a1d4d0c2f7a467dbbc0bfdf55bccf7 (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 摘要………………………………………………………………………………… I
Abstract………………………………………………………………………………II 目錄…………………………………………………………………………………III 圖次…………………………………………………………………………………VI 表次…………………………………………………………………………………VII 第一章 緒論………………………………………………………………………1 第二章 文獻回顧…………………………………………………………………4 第一節 鯉魚疱疹病毒的感染與控制…………………………………………4 1.1 鯉魚疱疹病毒的分類……………………………………………………4 1.2 傳播方式…………………………………………………………………5 1.3 臨床症狀…………………………………………………………………6 1.4 控制………………………………………………………………………7 第二節 魚類的免疫系統………………………………………………………10 2.1 非特異性免疫……………………………………………………………10 2.1.1 補體(Complement)………………………………………………10 2.1.2 凝集素(lectin)……………………………………………………12 2.1.3 干擾素(interferon)…………………………………………………13 2.1.4 殺手細胞(killer cell)與巨噬細胞(Macrophage)………………15 2.2 特異性免疫………………………………………………………………17 2.2.1 體液免疫(Humoral immunity)……………………………………17 2.2.2 細胞免疫(Cell-mediated immunity)………………………………19 第三節 魚類病毒疫苗成效與施用方法………………………………………20 3.1 疫苗現況…………………………………………………………………20 3.2 疫苗投予方法……………………………………………………………25 3.2.1 注射…………………………………………………………………25 3.2.2 浸泡…………………………………………………………………26 3.2.3 口服…………………………………………………………………26 第三章 材料與方法………………………………………………………………28 第一節 實驗設計與流程………………………………………………………28 第二節 製作去活化疫苗………………………………………………………29 2.1 病毒液製備………………………………………………………………29 2.2 病毒去活化………………………………………………………………30 2.3 DNA萃取與聚合酶鏈鎖反應(Polymerase chain reaction)…………31 2.4 巢式聚合酶鏈鎖反應(Nested-polymerase chain reaction)…………34 第三節 實驗魚接種疫苗………………………………………………………35 3.1 實驗材料…………………………………………………………………35 3.2 實驗方法…………………………………………………………………35 3.2.1 Naive 魚隻疫苗接種………………………………………………36 3.2.2 Carrier 魚隻疫苗接種………………………………………………37 3.2.3 魚隻含佐劑疫苗接種………………………………………………38 第四節 實驗魚攻毒試驗………………………………………………………40 4.1 實驗材料…………………………………………………………………40 4.2 實驗方法…………………………………………………………………40 第五節 養殖場魚隻疫苗接種…………………………………………………41 5.1 實驗材料…………………………………………………………………41 5.2 實驗方法…………………………………………………………………41 第六節 血清取得與血清中和試驗……………………………………………42 6.1 實驗材料…………………………………………………………………42 6.2 實驗方法…………………………………………………………………42 第七節 組織中病毒定量………………………………………………………43 7.1 實驗材料…………………………………………………………………43 7.2 實驗方法…………………………………………………………………43 第四章 結果………………………………………………………………………45 第一節 病毒液製備結果與福馬林去活病毒液隻細胞檢測…………………45 第二節 Naive組血清中和抗體力價與疫苗保護效力………………………45 第三節 Carrier組血清中和抗體力價與疫苗保護效力………………………46 第四節 攻毒後魚體組織中之病毒量…………………………………………46 第五節 不同免疫路徑之血清抗體力價比較…………………………………47 第六節 不同魚隻來源之血清抗體力價比較…………………………………47 第七節 Naive魚隻接種含佐劑隻疫苗後,其中和抗體力價、疫苗保護效力病毒訊號偵測…………………………………………………………………………47 第八節 Carrier魚隻接種含佐劑隻疫苗後,其中和抗體力價、疫苗保護效力病毒訊號偵測…………………………………………………………………………48 第九節 養殖場接種疫苗後血清中和力價變化情形…………………………49 第五章 討論………………………………………………………………………63 第一節 福馬林去活化…………………………………………………………63 第二節 死毒疫苗之接種路徑…………………………………………………64 第三節 抗體與病毒感染的關聯性……………………………………………67 第四節 佐劑對於死毒疫苗之影響……………………………………………70 第六章 結論………………………………………………………………………73 第七章 參考文獻…………………………………………………………………74 | |
| dc.language.iso | zh-TW | |
| dc.subject | 錦鯉?疹病毒 | zh_TW |
| dc.subject | 死毒疫苗 | zh_TW |
| dc.subject | 佐劑 | zh_TW |
| dc.subject | adjuvant | en |
| dc.subject | Koi Herpesvirus | en |
| dc.subject | Inactivated vaccine | en |
| dc.title | 評估台灣錦鯉疱疹病毒福馬林去活化疫苗引起之免疫效果與保護效力 | zh_TW |
| dc.title | Evaluate the immunogenicity and protective effects of formalin inactivated Taiwan Koi Herpesvirus vaccine | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張本恆(Pen-Heng Chang),齊肖琪(Shau-Chi Chi),涂堅(Chien Tu) | |
| dc.subject.keyword | 錦鯉?疹病毒,死毒疫苗,佐劑, | zh_TW |
| dc.subject.keyword | Koi Herpesvirus,Inactivated vaccine,adjuvant, | en |
| dc.relation.page | 85 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2014-08-15 | |
| 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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