臺灣博碩士論文加值系統

本論文為調查台灣地區國外輸入犬於隔離檢疫期間之腸道內寄生蟲感染情況及建立單包絛蟲糞內抗原診斷技術。研究進行期間於2004年1月至2005年6月，收集分別來自13個輸出國家的497個檢疫犬糞便檢體。檢查犬隻腸道內感染之蠕蟲及原蟲，使用的糞便檢查法包含直接塗抹法、糖水浮游法以及福馬林-乙酸乙酯沉澱法；鏡檢發現蠕蟲蟲卵及原蟲卵囊共計73例（14.7%）。為單一性感染檢體以犬球蟲出現的比例為最高5.8%（29/497），其次為鈎蟲4.8%（24/497），犬鞭蟲1.0%（5/497），犬蛔蟲0.6%（3/497）及犬小蛔蟲0.2%（1/497）。混合感染者11例（2.2%），混合感染者以犬鞭蟲與鈎蟲混合感染比例為最高1.4%（7/497），其次為犬蛔蟲與犬鞭蟲混合感染0.4%（2/497），犬球蟲與鈎蟲混合感染及犬蛔蟲與鈎蟲混合感染同為0.2%（1/497）。
包絛蟲（Echinococcus spp.）造成的包囊蟲症（hydatid disease; echinococcosis）是人畜共通性的重要寄生蟲性疾病，致病原為感染人類及草食性有蹄類動物之包絛蟲幼蟲，此種寄生蟲之終宿主主要為犬科動物，人類因食入犬隻終宿主糞便中之蟲卵而感染。單包絛蟲糞內抗原診斷技術開發，應用三明治酵素免疫分析法（sandwich ELISA）的原理，使用以單包絛蟲蟲體抗原免疫實驗兔之多株抗體，檢測攙入未感染蠕蟲犬隻糞便內之單包絛蟲抗原；並連同感染瓜實絛蟲、犬蛔蟲及鈎蟲的犬隻糞便檢體進行交叉反應試驗（cross-reactivity test），結果顯示包絛蟲與鈎蟲及犬蛔蟲無交叉反應，但與犬類常見的瓜實絛蟲發生交叉反應現象；研究中應用抗體吸附技術，確實排除與瓜實絛蟲間之交叉反應。以偵測糞內抗原為基礎的酵素免疫分析法應用於臨床診斷及檢疫將可提供快速且大量的篩選，並可提早篩檢出處於潛伏期或未排蟲卵之已感染動物。

The objectives of this study are to investigate the intestinal parasitic infections in imported dog by fecal examination, and also to develop the coproantigen sandwich ELISA to detect Echinococcus granulosus infection in the canine host. Feces of quarantined dogs collected between January, 2004 to June, 2005, in Taiwan, were examined by the direct smear method, Sheather''s sugar flotation method and formalin-ethyl acetate method for intestinal helminth and protozoan infection. Of the 497 dogs imported from 13 countries, 73 were found to be infected with at least one species of intestinal parasite. The parasites detected were oocysts of Isospora canis 5.8% (29/497) and eggs of hookworms 4.8% (24/497), Trichuris vulpis 1.0% (5/497), Toxocara canis 0.6% (3/497) and Toxascaris leonina 0.2% (1/497) in single infection. Dogs with two parasite species infection consist of 1.4%(7/497) for T. vulpis with hookwroms, 0.4% (2/497) for T. canis with T. vulpis, 0.2% (1/497) for I. canis with hookwroms, 0.2% (1/497) for T. canis with hookwroms, respectively. Echinococcosis is an important zoonotic parasitic disease. We developed a Sandwich ELISA using polyclonal antibodies raised in rabbits against the somatic antigen of E. granulosus to detect the coproantigen in the infected dog feces. No cross-reactivity was observed for feces of dogs with T. canis and Ancylostoma caninum infection. Cross-reactivity to D. caninum was eliminated by absorbing the polyclonal antibody with D. caninum tapeworm and then using them in the Sandwich ELISA.

中文摘要.......i
英文摘要.......ii
目 錄.......iii
表 次.......vi
圖 次.......vii
第一章 緒言.......1
第二章 文獻探討.......4
第一節 犬隻的輸入檢疫.......4
第二節 台灣地區流浪犬寄生蟲相關調查報告.......4
第三節 犬隻腸道內寄生蟲.......5
一、 絛蟲.......5
二、 蛔蟲.......5
三、 鈎蟲.......6
四、 鞭蟲.......7
五、 糞小桿線蟲.......7
六、 球蟲.......8
第四節 糞內抗原檢測犬隻單包絛蟲症之探討.......8
一、 包絛蟲的型態分類.......9
二、 單包絛蟲的分類.......10
三、 單包絛蟲的生活史.......12
四、 單包絛蟲之流行病學.......13
五、 人類之包囊蟲症.......15
第五節 糞內抗原檢測之原理與應用.......15
一、 糞內抗原檢測系統.......15
二、 糞內抗原之相關研究.......16
三、 動物感染試驗中蟲卵與糞內抗原檢出的時機差異.......16
四、 比較糞內抗原檢測與血清中抗體檢測之差異.......17
五、 糞內抗原的敏感度與特異性.......17
第三章 材料與方法.......19
第一節 輸入犬糞便檢體寄生蟲檢查.......19
一、 糞便檢體來源.......19
二、 顯微鏡鏡檢.......19
（一） 直接塗抹法.......19
（二） 糖水浮游法.......19
（三） 福馬林-乙酸乙酯沉澱法.......20
第二節 單包絛蟲糞內抗原診斷.......20
一、 單包絛蟲蟲體抗原的製備.......20
二、 蟲體抗原蛋白質濃度測定.......21
三、 抗單包絛蟲多株抗體製備.......21
（一） 實驗動物.......22
（二） 免疫計劃.......22
（三） 雙重免疫擴散法.......22
（四） 間接酵素結合免疫吸附法偵測之步驟.......23
（五） 純化濃縮多株抗體.......24
（六） 過氧化氫酶標定多株抗體.......25
四、 糞內抗原的製備.......26
五、 抗原攙入反應試驗.......26
六、 交叉反應試驗.......28
七、以瓜實絛蟲成蟲蟲體預先吸收第一抗體.......29
八、 偵測實驗感染犬之糞便內糞內抗原.......29
第四章 結果.......31
第五章 討論.......48
參考文獻.......51
附 件 一.......59
附 件 二.......60
附 錄.......67

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