劉振軒2006-07-262018-07-092006-07-262018-07-092001http://ntur.lib.ntu.edu.tw//handle/246246/28694犬瘟熱(canine distemper, CD) 是一種嚴重感染犬科動物，並具高度傳染性之病毒 性疾病。病原以侵害犬中樞神經、呼吸及消化系統為主。1995 年起，在台灣陸續出現成 犬疑似犬瘟熱引起脫髓鞘性腦炎(demyelinating encephalitis) 之病例。但是對於病原的 確認以病理學觀察仍嫌不足，因此，本研究係嘗試結合並比較形態病理學及反轉錄聚合鈳 鏈反應(RT-PCR) 診斷方法，以確認台灣地區之犬瘟熱病毒感染及其型式，同時測定 CDV 感染後腦脊髓液與腦組織中2´, 3´-cyclic nucleotide 3´-phosphohydrolase (CNP) 活性 之變化以評估做為脫髓鞘疾病診斷之可能性。自民國88 年3 月至90 年3 月間，由全省 各地區動物醫院及棄犬收容中心，收集33 隻疑似犬瘟熱症狀之患犬，進行病理學檢查、 免疫組織化學染色、組織化學染色及穿透式電子顯微鏡觀察形態病理學變化。同時，採 取新鮮中樞神經系統、淋巴結、肺臟、腦脊髓液及全血，使用RT-PCR 以偵測CD 病毒 磷酸蛋白基因(phosphoprotein gene)。結果發現16 例(16/33) 具有特徵性病理變化及出 現典型包涵體而診斷為CD;使用RT-PCR 診斷，則陽性病例增為27 例(27/33)。此外， 本研究亦發現2 例脫髓鞘性腦炎，經RT-PCR 產物之核酸定序分析結果，證實為CDV 感染所致。本實驗結合病理學及RT-PCR 核酸偵測技術，確認台灣地區有全身性及脫髓 鞘性犬瘟熱病例，RT-PCR 技術之應用，不僅提升診斷之特異性及敏感性，而且可以提 供為臨床診斷的實用工具。此外，利用分光光度法偵測CDV 感染後腦白質部組織及腦 脊髓液之CNP 活性，結果發現CDV 感染同時伴隨脫髓鞘病變的犬隻，腦脊髓液中之 CNP 活性呈現明顯上升(p<0.05);而CDV 感染的犬隻，無論有無出現脫髓鞘病變，腦 組織中之CNP 活性皆明顯下降(p<0.05)。由此推論，此項CNP 活性測定技術的建立， 未來有潛力應用於評估中樞神經系統髓鞘的流失情形，而當做脫髓鞘性疾病的輔助診斷 工具。Canine distemper (CD) is a highly contagious viral disease, which has been reported in dogs for many years in Taiwan. Ever since 1995, apart from systemic CD, there has been continuous emergence of CD-like demyelinating encephalitis in adult dogs. The goal of this study is to identify CD in Taiwan using pathology and RT-PCR as diagnostic tools and to assay 2´, 3´-cyclic nucleotide 3´-phosphohydrolase (CNP) activity of cerebrospinal fluid and brain tissue with canine distemper virus infection. This report collected 33 cases from animal hospitals throughout the country as well as from the dog pounds. The affected dogs died either natural cause or by euthanasia. All cases were conducted to perform pathological investigation, ABC immunohistochemistry, luxol-fast blue cresyl echt violet (LFB-CEV) histochemistry as well as transmissible electron microscopy. Phosphoprotein gene of canine distemper virus (CDV) was used as the target gene for RT-PCR. Under histopathological examination, 16 cases (48.48%) were diagnosed as CD. With the aid of RT-PCR, 27 cases (81.82%) were diagnosed as CDV positive. This study also found that 2 cases of demyelinating encephalitis were diagnosed as CDV infection with histopathology, LEB-CEV stain and RT-PCR. On conclusion, both systemic and demyelinating CD were identified in Taiwan using RT-PCR along with histopathology. Combination of the two techniques not only raises CD diagnostic rate, but also provides a more rapid, specific, and sensitive method to confirm CDV infection. Additionally, spectrophotography was employed in the CNP assay of the brain tissues and cerebrospinal fluid. The results showed that CNP activity in CD infected dogs with demyelinating lesion had a significant elevation in the CSF (p<0.05). On the other hand, CD infected dogs with or without demyelinating lesion demonstrated a significant decline in CNP activity (p<0.05). Based on the results, CNP assay will be potentially to reflect status of myelin loss and diagnose demyelinating diseases.application/pdf59701 bytesapplication/pdfzh-TW國立臺灣大學獸醫學系暨研究所犬瘟熱脫髓鞘性腦炎反轉錄聚合鏈鎖反應canine distemperdemyelinating encephalitisRT-PCRCNP assayreporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/28694/1/892313B002196.pdf