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研究生:陳昱憲
研究生(外文):Yu-Shian Chen
論文名稱:以巢式聚合酶連鎖反應調查台灣家貓血液寄生蟲之感染疫情
論文名稱(外文):Detection of the Blood Parasite Infections in Domestic Cats from Taiwan by Nested Polymerase Chain Reaction
指導教授:曾秋隆曾秋隆引用關係
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
畢業學年度:96
語文別:中文
論文頁數:59
中文關鍵詞:犬型艾利希體貓血巴東體
外文關鍵詞:Ehrlichia canisMycoplasma haemofelis
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近年來,台灣地區作為伴侶動物的貓隻數目日益漸增。在小動物臨床醫學上,以往許多只見於在犬隻身上發生之血液寄生蟲感染症,如犬型艾利希體症、犬焦蟲症…等,近年來都有在貓隻上發現零星的病例報告。本實驗的目標是利用巢式聚合酶連鎖反應以及核酸定序對小動物臨床上常見的血液寄生蟲做疫情調查。以gp36基因做為檢測犬型艾利希體 (Ehrlichia canis) 之標的、gltA基因做為檢測血小板型艾利希體 (Anaplasma platys)之標的以及16S rRNA做為檢測貓血巴東體 (Mycoplasma haemofelis以及Candidatus Mycoplasma haemominutum) 之標的。從兩百個家貓血液檢體中,一共篩檢到41個陽性病歷 (20.5 %),其中包含Ehrlichia canis感染11例 (5.5 %)、Anaplasma platys感染4例 (2.0 %)、、Mycoplasma haemofelis感染4例 (2.0 %) 以及Candidatus Mycoplasma haemominutum感染25例 (12.5 %)。至於病原的傳播路徑,除了以往認知中體外寄生蟲媒介性傳播外,可以合理的推測如貓隻間打鬥導致帶菌血液從傷口直接侵犯性傳播或是媒介間病原的傳播對於這些疾病在流行病學上可能都佔有一定重要性的地位。
The population of cat for companion animals get a huge increasing in Taiwan recently years. Some blood parasite infection diseases used to observed on dogs, like Ehrlichia canis、Babesia canis…etc, have lots of case reports on cats. The aim of this study will try to detect the prevalence of blood parasite infections used to observed in clinical veterinary, by using the combination of nested polymerase chain reaction (nPCR) and nucleotide sequencing assays to determine the prevalence of Ehrlichia canis (Ec) with gp36 gene、Anaplasma platys (Ap; formerly E. platys) with gltA gene、Mycoplasma haemofelis (Mhf) and Candidatus Mycoplasma haemominutum (Mhm) with 16S rRNA in domestic cats in Taiwan. DNA from one or more of the organisms was amplified from 41 of 200 blood samples (28.0%). DNA consistent with Ec 11 (5.5 %), Ap 4 (2.0 %), Mhf 4 (2.0 %) and Mhm 25 (12.5 %) were detected. Besides an ostensible vector-brone transmission, direct transmission by aggressive interaction of cats or interspecies transmission might play a role in epidemiology of these organisms.
中文摘要…………………………………………………………………I
英文摘要………………………………………………………………II
目次……………………………………………………………………III
中英對照表……………………………………………………………VII
圖次……………………………………………………………………VIII
表次………………………………………………………………………IX
第一章 緒言…………………………………………………………1
第二章 文獻探討……………………………………………………3
2-1 艾利希體症
2-1.1 艾利希體症之歷史背景……………………………………3
2-1.2 艾利希體症之研究背景及目的……………………………5
2-1.3 病原體………………………………………………………5
2-1.4 分類…………………………………………………………6
2-1.5 傳播途徑……………………………………………………7
2-1.6 臨床症狀……………………………………………………7
2-1.7 診斷方法……………………………………………………8
2-2 血巴東體症
2-2.1 血巴東體症之歷史背景……………………………………11
2-2.2 血巴東體症之研究背景及目的……………………………11
2-2.3 病原體………………………………………………………12
2-2.4 分類…………………………………………………………12
2-2.5 傳播途徑……………………………………………………13
2-2.6 臨床症狀……………………………………………………13
2-2.7 診斷方法……………………………………………………13
第三章 材料與方法…………………………………………………16
3-1 檢體之取得………………………………………………………16
3-1.1 血液檢體之來源……………………………………………16
3-1.2 血液抹片之製作……………………………………………16
3-2 檢體 (貓隻血液) 基因體核酸之萃取…………………………16
3-3 聚合酶連鎖反應及巢式聚合酶連鎖反應………………………17
3-4 引子之設計………………………………………………………17
3-4.1 Ehrlichia canis gp36基因………………………………17
3-4.2 Ehrlichia platys gltA基因………………………………18
3-4.3 Mycoplasma haemofelis 16S rRNA基因…………………18
3-5 聚合酶連鎖反應產物之確認……………………………………19
3-6 聚合酶連鎖反應產物之選殖……………………………………20
3-6.1聚合酶連鎖反應產物之純化…………………………………20
3-6.2聚合酶連鎖反應產物之選殖…………………………………20
3-7 質體DNA之純化…………………………………………………21
3-8 選殖產物之確認…………………………………………………21
3-9 聚合酶連鎖反應產物定序以及序列分析………………………22
第四章 結果…………………………………………………………25
4-1 犬型艾利希體在貓隻感染情況…………………………………25
4-1.1 聚合酶連鎖反應之產物的確認………………………………25
4-1.2 以聚合酶連鎖反應檢測貓隻感染Ehrlichia canis結果…25
4-1.3 Ehrlichia canis定序確認…………………………………25
4-1.4 貓隻感染犬型艾利希體臨床症狀之觀察……………………26
4-1.5 Ehrlichia canis引子專一性測試…………………………26
4-1.6 Ehrlichia canis引子敏感度測試…………………………26
4-2 血小板型艾利希體在貓隻感染情況……………………………30
4-2.1 聚合酶連鎖反應反應產物之確認……………………………30
4-2.2 以聚合酶連鎖反應檢測貓隻感染Ehrlichia platys結果…30
4-2.3 Anaplasma platys定序確認…………………………………30
4-2.4 貓隻感染血小板型艾利希體臨床症狀之觀察………………30
4-2.5 Anaplasma platys引子專一性測試…………………………31
4-2.6 Anaplasma platys引子敏感度測試…………………………31
4-3 貓血巴東體在貓隻感染情況……………………………………35
4-3.1 聚合酶連鎖反應之產物的確認………………………………35
4-3.2 以聚合酶連鎖反應檢測貓隻感染Mycoplasma haemofelis
結果……………………………………………………………35
4-3.3 Mycoplasma haemofelis定序確認…………………………35
4-3.4 貓隻感染血巴東體臨床症狀之觀察…………………………36
第五章 討論…………………………………………………………45
參考文獻…………………………………………………………………52
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