闕玲玲臺灣大學：獸醫學研究所陳佩琦Chen, Pey-ChyiPey-ChyiChen2007-11-282018-07-092007-11-282018-07-092005http://ntur.lib.ntu.edu.tw//handle/246246/59989貓冠狀病毒(feline coronavirus)感染貓的臨床表現可分為腸道型及腹膜炎型。本實驗係調查台北市私人動物醫院及台灣大學附設動物醫院，臨床就診懷疑有貓冠狀病毒感染的貓隻臨床檢體，配合巢式反轉錄聚合酶鏈鎖反應(RT-nPCR)針對貓冠狀病毒高保留之3’UTR區域進行篩選，並針對由台灣大學附設動物醫院自2000年六月至2004年四月，共40隻自臨床檢體偵測到病毒核酸並診斷為滲出性貓傳染性腹膜炎(feline infectious peritonitis, FIP)而死亡的病例進行分析。發現其中腹水及胸水樣本的貓冠狀病毒檢測陽性率皆為100%。此外，陽性病例中品種的比例以純種品系佔多數，年齡分布主要為3月齡至3歲齡(30/40, 75%)，另外分析其中20隻患貓的血液學及血清生化學方面，貧血的現象佔70%(14/20)、淋巴球減少的情形佔90%(18/20)、而有70%(14/20)患貓的A/G ratio<0.7。本研究嘗試進行病毒分離及血清學鑑定分析，發現樣本中以血清型第一型居多。同時分析一臨床陽性滲出液樣本中病毒7a/7b基因靠近3’端的475 bp，與國外序列進行比對，發現此區域與美國的分離株的FIPV NOR-15株最為接近，具有95.4%的相似度。Feline coronavirus (FCoV) infection in cats may cause peritonitis (FIP) or a milder form of enteritis. The study is to investigate cats from private clinic and National Taiwan University Animal Hospital, with correlation of a nested reverse transcription polymerase chain reaction (RT-nPCR) assay to detect feline coronavirus highly conserved 3’ untranslated region in clinical specimens. A total of 40 cats from June 2000 to April 2004 were screened to be feline coronavirus positive and died with the diagnosis as effusive FIP . Among these 40 cats, FCoV can be detected in all of the effusion specimens. Most of the cats were purebred, with 75% (30/40)aged from 3 month to 3 year. Among them 20 with blood test data were analysed, 70% (14/20)appeared to have anemia, 90% (18/20)with lymphopenia and 70% (14/20) with A/G ratio lower than 0.7. Virus isolation were attempted using body effusion collected from positively diagnosed animals. Limited amount of coronavirus can only be detected up to the third passage by RT-nPCR. Serologic assay revealed that FCoV type I might be predominant in Taipei area. Sequence comparison were made from 475bp of 7a/7b gene, and the result shows high degree of homology to several other known FCoV strains, including FIPV strain NOR-15 isolated in the USA, which are 95.4% identical in this region.目錄 目錄............................................................................................................................... ii 圖次................................................................................................................................v 表次...............................................................................................................................vi 摘要............................................................................................................................. vii Abstract..................................................................................................................... viii 第一章、文獻探討.......................................................................................................1 第一節、貓冠狀病毒感染之歷史背景...................................................................................... 1 1.1 貓傳染性腹膜炎...................................................................................................................................... 1 1.2 貓腸道型冠狀病毒感染.......................................................................................................................... 2 第二節、貓冠狀病毒之宿主範圍.............................................................................................. 2 第三節、貓冠狀病毒之結構與生化特性.................................................................................. 3 3.1 冠狀病毒科(family Coronaviridae) .........................................................................................................3 3.2 貓冠狀病毒結構與特性.......................................................................................................................... 6 第四節、貓冠狀病毒感染之流行病學...................................................................................... 7 第五節、致病機轉...................................................................................................................... 8 第六節、臨床症狀.................................................................................................................... 10 第七節、貓冠狀病毒感染之診斷............................................................................................ 11 7.1 臨床病理學檢查................................................................................................................................... 12 7.2 實驗室診斷........................................................................................................................................... 12 7.3 病理學檢查........................................................................................................................................... 13 第八節、貓冠狀病毒之疫苗發展............................................................................................ 13 第二章、緒言.............................................................................................................16 第三章、材料與方法.................................................................................................17 第一節、實驗設計.................................................................................................................... 17 第二節、實驗材料及方法........................................................................................................ 17 2.1 病材的收集與記錄................................................................................................................................ 17 2.2 貓冠狀病毒RNA 萃取及RT-PCR ....................................................................................................... 18 2.3 貓冠狀病毒7a/7b gene 序列分析及比對.............................................................................................. 19 2.4 貓冠狀病毒的分離及檢測.................................................................................................................... 20 2.5 以FA substrate slide 分析貓冠狀病毒血清型別.................................................................................. 21 第四章、結果.............................................................................................................24 第一節、病材的收集與記錄.................................................................................................... 24 第二節、貓冠狀病毒7a/7b gene 序列分析............................................................................ 26 第三節、貓冠狀病毒的分離及檢測........................................................................................ 27 第四節、以FA substrate slide 分析貓冠狀病毒血清型別..................................................... 27 第五章、討論.............................................................................................................46 附錄一、濕式FIP 發病貓隻之血液學及血清生化學數值.....................................51 附錄二、所用儀器廠商名稱.....................................................................................53 附錄三、病史及臨床資料分析記錄表.....................................................................54 附錄四、主要溶液的配製.........................................................................................55 附錄五、濕式FIP 發病貓以及非FIP 就診貓隻血液學及血清生化學 數值之比較..................................................................................................................57 參考文獻......................................................................................................................58en-US貓傳染性腹膜炎貓冠狀病毒feline infectious peritonitisfeline coronavirusthesis