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| 研究生： |
許家福 Hsu, Chia-Fu |
|---|---|
| 論文名稱： |
豬生殖與呼吸綜合症病毒活毒減毒疫苗搭配不同佐劑之效力評估 Efficacy of Porcine Reproductive and Respiratory Syndrome Virus Live Attenuated Vaccine with Different Adjuvants |
| 指導教授： |
邱明堂
Chiou, Ming-Tang 林昭男 Lin, Chao-Nan |
| 學位類別： |
碩士 Master |
| 系所名稱： |
獸醫學院 - 獸醫學系所 Department of Veterinary Medicine |
| 畢業學年度： | 108 |
| 語文別： | 中文 |
| 論文頁數： | 91 |
| 中文關鍵詞： | 豬生殖與呼吸綜合症 、活毒減毒疫苗 、佐劑 、免疫反應 |
| 外文關鍵詞： | porcine reproductive and respiratory syndrome, live attenuated vaccine, adjuvants, immune response |
| DOI URL： | http://doi.org/10.6346/NPUST202000231 |
| 相關次數： | 點閱：38 下載：0 |
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中文摘要 I
Abstract III
謝 誌 V
目 錄 VI
圖表目錄 IX
第 1 章 前 言 1
第 2 章 文獻回顧 4
2.1豬生殖與呼吸綜合症病毒 4
2.1.1疾病爆發與經濟損失 4
2.1.2病毒介紹 5
2.1.2.1病毒學及命名 5
2.1.2.2基因體構造及表現 6
2.1.3感染宿主 7
2.1.4傳播途徑 7
2.1.5臨床症狀 8
2.1.6 PRRSV感染後之反應 9
2.1.7 PRRSV對先天性免疫反應之影響 9
2.1.7.1先天性免疫反應對後天性免疫反應之影響 10
2.1.8 PRRSV對後天性免疫反應之影響 10
2.1.8.1體液性免疫反應 10
2.1.8.2抗體依賴性增強 11
2.1.8.3中和抗體之保護力 11
2.1.8.4細胞性免疫反應 12
2.1.9 PRRSV躲避宿主免疫反應之機制 13
2.1.9.1先天性免疫反應 13
2.1.9.2後天性免疫反應 14
2.1.9.2.1體液性免疫反應 14
2.1.9.2.1.1中和抗體決定位之醣基化 14
2.1.9.2.1.2誘騙表位之出現 14
2.1.9.2.2細胞性免疫反應 15
2.1.10疾病預防與控制 15
2.1.11疫苗開發之困境 16
2.1.12 理想之PRRSV疫苗 16
2.2佐劑 (adjuvants) 17
2.2.1背景與歷史 17
2.2.2種類與功能 18
2.2.2.1 CpG ODNs 18
2.2.2.2介白素 (Interleukin, IL) 19
第 3 章 材料與方法 20
3.1 實驗設計 20
3.2.1 細胞存活試驗 (佐劑毒性試驗) 21
3.2.2 病毒感染試驗 21
3.2.2.1 核酸萃取 22
3.2.2.2 即時定量聚合酶鏈鎖反應 22
3.3 豬生殖與呼吸綜合症病毒與佐劑 22
3.3.1 田間野外病毒分離 23
3.3.2 病毒純化 23
3.3.3 病毒力價測定 24
3.4 動物實驗 24
3.4.1 實驗動物 24
3.4.2 實驗流程 24
3.4.3 血液樣本採集 25
3.4.4 實驗動物犧牲 25
3.5 周邊血液單核細胞分離 25
3.5.1 IFN-γ ELISpot測定 26
3.5.2 PRRS特異性PBMCs增殖試驗 26
3.6 實驗動物肺臟肉眼及顯微評分 27
3.7 原位雜和反應 (In situ hybridization, ISH) 27
3.7.1 探針之合成 27
3.7.2 原位雜合染色 28
3.8 血清學檢測 28
3.8.1 酵素連結免疫分析法 28
3.8.2 病毒中和抗體試驗 29
3.8.2.1 間接免疫螢光分析法 29
3.8.3 病毒血症檢測 30
3.8.4 血清IL-6及TNF-α檢測 30
3.9 統計分析與圖表繪製 30
第 4 章 結 果 31
4.1動物實驗前試驗 31
4.1.1細胞存活試驗結果 31
4.1.2病毒感染試驗 31
4.2 PRRSV感染PAM之細胞病變作用 31
4.3動物實驗結果 32
4.3.1實驗動物之體溫變化 32
4.3.1.1 PRRSV MLV疫苗免疫後之體溫變化 32
4.3.1.2 PRRSV野外分離株攻毒之體溫變化 32
4.3.2實驗動物之存活率 32
4.3.2.1死亡豬隻病理學診斷 33
4.3.3 實驗動物血清特異性PRRSV抗體 33
4.3.4 實驗動物血清PRRSV病毒量變化 34
4.3.5 PRRSV中和抗體結果 34
4.3.6 IFN-γ ELISpot測定結果 34
4.3.7 PRRSV特異性PBMCs增殖試驗結果 35
4.3.8 實驗動物之肺臟評分 35
4.3.9 原位雜和反應結果 35
第 5 章 討 論 37
參考文獻 65
附錄 89
作者簡介 91
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