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研究生:郭忠融
研究生(外文):Chung-Zung Kuo
論文名稱:台灣乳牛感染牛呼吸道融合病毒之調查
論文名稱(外文):Survey of bovine respiratory syncytial virus infection in dairy cattle in Taiwan
指導教授:莊士德莊士德引用關係
指導教授(外文):Shih-Te Chuang
口試委員:陳鵬文葉光勝
口試委員(外文):Jacky Peng-Wen ChanKuang-Sheng Yeh
口試日期:2015-07-21
學位類別:碩士
校院名稱:國立中興大學
系所名稱:獸醫學系暨研究所
學門:獸醫學門
學類:獸醫學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:51
中文關鍵詞:牛呼吸道融合病毒
外文關鍵詞:bovine respiratory syncytial virus
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自1970年代起,牛呼吸道融合病毒(bovine respiratory syncytial virus, BRSV)就成為造成乳牛呼吸道疾病的重要病原之一,2001年蔡發表首篇關於台灣BRSV的盛行率報告,顯示BRSV已經普遍存在於台灣乳牛場之中,但近年來並無關於BRSV的盛行率報告。本研究的目的在於調查台灣BRSV的盛行率及偵測BRSV的抗原。在2014年間,從15個縣市84個牧場中蒐集了1192頭牛隻血清來進行ELISA抗體檢測;另外,自2012年10月至2014年9月間蒐集的387頭牛隻的鼻腔拭子和血液樣本,利用nested RT-PCR檢測BRSV抗原。總牛隻個體盛行率為43.4% (517/1192),場盛行率為79.8% (67/84),場內盛行率中位數為43.4%,範圍從0%至100%。將台灣劃作北、中、南和東四個區域,牛隻個體盛行率由高至低依序為南部(58.5%)、中部(55.1%)、北部(16.5%),最後是東部(12.0%),場盛行率由高至低依序為南部(95.0%)、中部(91.4%)、東部(58.3%),最後是北部(52.9%),各區域的個體和場盛行率在統計學上有顯著差異(p˂0.05)。此外,以nested RT-PCR檢測抗原,陽性率為1.6% (6/387)。本次調查結果顯示在台灣的乳牛場中普遍存在BRSV的感染,且呈現中南部BRSV的盛行率高於北部、東部。

Bovine respiratory syncytial virus (BRSV) is an important pathogen that causes bovine respiratory disease in dairy cattle since 1970s. According to Tsai’s report in 2001 which was the first report about the prevalence of BRSV in Taiwan, BRSV infection was a common respiratory disease in dairy herd in Taiwan. However, there was no report about the prevalence of BRSV infection recently. The aims of this study were to investigate the prevalence and to detect the antigen of BRSV in Taiwan. A total of 1192 bovine sera, in which the antibody of BRSV was detected by ELISA, were collected from 84 dairy herds in 15 counties in 2014. Additionally, nasal swabs and blood samples of 387 cows, in which the antigen of BRSV was detected by nested RT-PCR, were collected from October, 2012 to September, 2014. The prevalence of BRSV was 43.4% (517/1192), and 67 of 84 (79.8%) herds were examined positive. The intra-herd prevalence ranged between 0% and 100% (median=43.4%). Regional individual prevalence was highest in South (58.5%) than in Center (55.1%), North (16.5%), and East (12.0%). Regional herd prevalence was highest in South (95.0%) than in Center (91.4%), East (58.3%), and North (52.9%). Individual and herd prevalence were significantly different in region (p˂0.05). The positive rate of detection antigen of BRSV was 1.6% (6/387). In conclusion, BRSV infection is a common respiratory disease in dairy herd in Taiwan, and the prevalence in Center and South is higher than in North and East.

中文摘要…………………………………………………………………………… i
英文摘要…………………………………………………………………………… ii
目次…………………………………………………………………………............ iii
表圖目次……………………………………………………………….................... vi
第一章 前言…………………………………………………………………........ 1
第二章 文獻探討………………………………………………………………… 2
第一節 病毒名稱與歷史………………………………………………........ 2
第二節 流行病學………………………………………………………........ 2
一、 國外BRSV調查結果…………………………………………........ 2
(一) 國外血清學調查結果……………………………………........ 2
(二) 其他危險因子分析………………………………………........ 3
二、 台灣BRSV調查結果…………………………………………........ 4
第三節 病原特性與致病機制…………………………………………........ 4
第四節 傳播方式………………………………………………………........ 5
第五節 臨床症狀和病理變化…………………………………………........ 5
第六節 診斷方式………………………………………………………........ 6
一、 病毒抗原檢測…………………………………………………........ 6
(一) 病毒分離…………………………………………………........ 6
(二) 直接免疫螢光染色法或免疫組織化學染色法…………........ 7
(三) 反轉錄聚合酶鏈鎖反應…………………………………........ 7
二、 病毒抗體檢測…………………………………………………........ 8
(一) 病毒中和試驗……………………………………………........ 8
(二) 酵素連結免疫吸附法……………………………………........ 8
第七節 治療與預防……………………………………………………........ 9
一、 治療……………………………………………………………........ 9
二、 預防……………………………………………………………........ 10
第八節 BEF之基本介紹………………………………………………........ 11
第三章 材料與方法…………………………………………………………….... 15
第一節 BRSV盛行率之試驗動物選取與樣本處理………………………. 15
一、 血清樣本數估計…………………………………………………… 15
二、 血清樣本之蒐集…………………………………………………… 15
第二節 偵測BRSV抗原樣本之初步處理………………………………… 15
一、 小牛樣本…………………………………………………………… 15
(一) 鼻腔拭子……………………………………………………… 15
(二) 全血樣本……………………………………………………… 15
(三) 臟器組織……………………………………………………… 16
二、 成牛樣本…………………………………………………………… 16
(一) 鼻腔拭子……………………………………………………… 16
(二) 全血樣本……………………………………………………… 16
(三) 臟器組織……………………………………………………… 16
第三節 以酵素連結免疫吸附法檢測BRSV抗體………………………… 17
一、 實驗之標準操作參照檢驗套組說明……………………………… 17
二、 結果判讀方式……………………………………………………… 17
第四節 巢式反轉錄聚合酶鏈鎖反應(nested RT-PCR)……………………. 18
一、 RNA萃取…………………………………………………………... 18
二、 cDNA合成…………………………………………………………. 19
三、 Nested RT-PCR引子對序列……………………………………….. 19
(一) BRSV………………………………………………………….. 19
(二) BEF……………………………………………………………. 19
四、 Nested RT-PCR增幅反應………………………………………….. 20
(一) BRSV………………………………………………………….. 20
(二) BEF……………………………………………………………. 20
五、 電泳分析…………………………………………………………… 21
六、 定序及分析………………………………………………………… 21
第五節 統計分析…………………………………………………………… 21
第四章 結果……………………………………………………………………… 22
第一節 蒐集樣本…………………………………………………………… 22
一、 成牛血清樣本……………………………………………………… 22
二、 小牛樣本…………………………………………………………… 22
三、 成牛樣本…………………………………………………………… 23
第二節 BRSV盛行率結果………………………………………….… 23
一、 台灣總盛行率結果………………………………………………… 23
二、 各縣市盛行率結果………………………………………………… 23
三、 各區域盛行率結果………………………………………………… 24
四、 各採樣時間盛行率結果…………………………………………… 24
第三節 BRSV抗原調查結果………………………………………………. 24
第四節 BEF病毒抗原調查結果與BRSV抗原調查結果之相關性比較…. 25
第五章 討論……………………………………………………………………… 30
第一節 BRSV盛行率調查…………………………………………………. 30
第二節 BRSV抗原偵測……………………………………………………. 33
第三節 BEF病毒抗原調查結果與BRSV抗原調查結果之相關性比較…. 34
第六章 總結……………………………………………………………………… 36
參考文獻…………………………………………………………………………… 37
附錄………………………………………………………………………………… 50
附錄一 台灣乳牛密度前20名地區….………………………………………… 50
附錄二 台灣乳牛場密度前20名地區…………………………………………. 51


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