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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 蔡向榮(Hsiang-Jung Tsai) | |
| dc.contributor.author | Ching-Hui Kuo | en |
| dc.contributor.author | 郭靜蕙 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:14:48Z | - |
| dc.date.copyright | 2010-08-16 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-10 | |
| dc.identifier.citation | 王秋文。台灣火雞鼻氣管鳥桿菌之生長特性、抗微生物劑感受性與巢氏聚合酶鏈鎖反應檢測之研究。嘉義大學獸醫學研究所碩士論文。嘉義。2007。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22277 | - |
| dc.description.abstract | 為了解台灣地區鳥鼻氣管桿菌(Ornithobacterium rhinotracheale)菌株對於奎諾酮類抗菌劑與gyrA基因之關係，所選用之試驗菌株來源為國立台灣大學獸醫學系預防醫學研究室所保存於2004 ~ 2006年間針對台灣地區屠宰場及禽鳥養殖場所採樣分離之O. rhinotracheale分離株，涵蓋物種包括雞、火雞、鴿、鴕鳥、鵪鶉、鳳頭蒼鷹等經濟動物及人工飼養寵物鳥共86株，以瓊脂稀釋法分別進行nalidixic acid、norfloxacin、ofloxacin、enrofloxacin與ciprofloxacin之奎諾酮類抗菌劑感受性試驗，結果O. rhinotracheale對上述抗菌劑的最小抑制濃度範圍分別為： 2~ > 128 μg/mL、< 0.125~ > 64μg/mL、< 0.125~ >32μg/mL、< 0.125~ > 8μg/mL及< 0.125~ > 16μg/mL。試驗同時亦針對分離株之DNA旋轉酶 (DNA gyrase- gyrA gene)進行序列分析，序列比對結果發現共有62株對奎諾酮類抗菌劑具有抗性，於GyrA第83或/與第87個胺基酸發生變異，其中85.5％ （53/62）及4.8％ （3/62）菌株於第83個胺基酸由絲胺酸（serine）分別突變為酪胺酸（tyrosin）及苯丙氨酸 (phenylalanine)； 9.8％（6/62）分別於第83及87個胺基酸由絲胺酸（serine）及天門冬氨酸（aspartic acid）突變為苯丙氨酸 (phenylalanine)及天門冬醯胺（aspargine）(5/6) 或丙氨酸 (alanine) (1/6)，且62株O. rhinotracheale之奎諾酮類抗菌劑感受性試驗結果同時對nalidixic acid （≧32 μg/mL； 96.8％）與norfloxacin （≧16 μg/mL； 90.3％）具有抗性，推測當O. rhinotracheale之GyrA第83或/與第87個胺基酸發生變異時與本菌對奎諾酮類抗菌劑產生之抗性有關（p＜0.01）。進一步利用高解析熱熔解分析方法成功的將四種不同型式之變異菌株以不同之熔解曲線區分開來，建立出快速鑑別O. rhinotracheale分離株之奎諾酮類抗藥性基因之診斷工具。另比較脈衝式電泳圖譜與抗菌劑感受性結果發現：於同時間地點與同動物來源所分離之菌株並非全然屬於同一基因型，推測可能因宿主之感染源複雜、禽場間之用藥方式不同、產生抗藥性的菌株因染色體DNA發生變異或菌株之基因體於Sal I限制酶切位上有些許差異而使PFGE圖譜有不同的基因型產生。 | zh_TW |
| dc.description.abstract | A total of 86 isolates of Ornithobacterium rhinotracheale were obtained from veterinary preventive medicine of National Taiwan University. Species include chickens, turkeys, pigeons, ostrichs, quails, and Accipiter trivirgatus respectively from 2004 to 2006. This study purpose is to analyze the quinolone resistant mutations of gyrA gene of O. rhinotracheale in Taiwan. Sixty-two (72.1 %, 62/86) quinolone resistant O. rhinotracheale strains were found by agar dilution method with the antibiotics nalidixic acid, norfloxacin, ofloxacin, enrofloxacin and ciprofloxacin ( the MIC range was 2 ~ > 128 μg/mL, < 0.125 ~ > 64 μg/mL, < 0.125~ > 32 μg/mL, <0.125 ~ > 8 μg/mL and < 0.125 ~ > 16 μg/mL, respectively) and sequencing of the polymerase chain reaction amplified products of the quinolone resistant determining regions (QRDRs) of gyrA showed that all isolates carrying single or double mutations in gyrA at codon 83 and 87. The mutation at 83 of GyrA from serine to tyrosine (S83T) and phenylalanine (S83P) was present in 85.5 %（53/62） and 4.8 %（3/62）, respectively. The double mutation at 83 and 87 of GyrA from serine and aspartic acid to phenylalanine and aspargine (5/6) or alanine (1/6) was present in 9.8 %（6/62）. The present study indicated that the presence of high-level resistance to quinolones in 62 O. rhinotracheale isolates (resistant to nalidixic acid and norfloxacin were ≧32 μg/mL; 96.8 % and ≧16 μg/mL; 90.3 %, respectively) was associated with mutations at codon 83 and 87 in GyrA (p <0.01). Sequence variants of four types were also revealed by high resolution melting (HRM) analytical scheme and established the rapid differentiation of gyrA mutations causing quinolone resistance in O. rhinotracheale. The compartment of pulsed-field gel electrophoresis (PFGE) results and susceptibility profile patterns showed that the strains isolated from the same location and species were not belonging to the same genotype. The resistance of the individual and the different antimicrobial disposal might cause the chromosomal variation on the Sal I restriction site of the quinolone resistant strains. And this resulted in the different genotypes in PFGE. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:14:48Z (GMT). No. of bitstreams: 1 ntu-99-R97629035-1.pdf: 1324790 bytes, checksum: 158e157268074ae4c86dc1a90486defd (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 口試委員審定書------------------ I
致謝--------------------II 中文摘要------------------------III 英文摘要------------------------V 目錄--------------------------VII 圖目錄------------------------XI 表目錄--------------------------XII 第一章 緒言---------------------1 第二章 文獻探討------------------2 第一節 鳥鼻氣管桿菌 (Ornithobacterium rhinotracheale)----2 1.1 歷史回顧--------------------2 1.2 分類與命名-----------------3 1.3 病原特性--------------------3 1.3.1 形態學-----------------3 1.3.2 生化性狀--------------4 1.4 感染途徑--------------------4 1.5 臨床症狀--------------------5 1.6 病理學變化-----------------5 1.6.1 肉眼病變-------------5 1.6.2 組織病理學病變--------------6 1.7 病原分離及鑑定--------------6 1.7.1 生化鑑定-------------7 1.7.1.1 API 20NE系統----------7 1.7.1.2 API-ZYM系統----------8 1.7.2 血清學檢測------------8 1.7.3 免疫組織化學染色--------------9 1.7.4 免疫螢光染色-----------------9 1.7.5 聚合酶鏈鎖反應----------------10 1.8 治療與控制----------------10 1.9 疫苗防治-------------------11 第二節 奎諾酮類抗菌劑 (Quinolone)------------13 2.1 Quinolone之歷史------------13 2.2 Quinolone之作用機制----------------14 2.3 Quinolone之藥物動力學----------------14 第三節 O. rhinotracheale之抗菌劑感受性試驗----------15 3.1 世界各國O. rhinotracheale抗菌劑感受性報告-------16 3.2 O. rhinotracheale之抗藥性機制----------19 第四節 O. rhinotracheale於分子生物學分型研究----------21 第五節 脈衝式電泳 (Pulsed-field gel electrophoresis, PFGE)----------23 第六節 高解析熱熔解分析 (High-resolution melting analysis, HRM) -----------------25 第三章 材料方法-----------------27 第一節 菌株來源與鑑定------------27 1.1 菌株來源-------------------27 1.2 菌株鑑定-------------------27 1.2.1 生化鑑定------------27 1.2.2 聚合酶鏈鎖反應鑑定----------28 1.2.2.1 核酸之萃取------------28 1.2.2.2 引子之選用------------29 1.2.2.3 聚合酶鏈鎖反應-------------29 1.2.2.4 瓊脂凝膠電泳分析-------------30 第二節 抗菌劑感受性試驗------------30 第三節 O. rhinotracheale DNA旋轉酶 (DNA gyrase- gyrA gene)之偵測-----------------31 第四節 O. rhinotracheale拓撲異構酶 (Toposiomerase IV parC gene)之偵測---------------------32 第五節 高解析熱熔解分析------------------33 第六節 脈衝式電泳試驗------------------34 6.1 菌塊包埋及清洗------------34 6.2 限制酶作用與脈衝電泳分析----------35 第七節 統計與分析-----------------35 7.1 菌株抗藥性百分率統計分析----------35 7.2 高解析熱熔解分析------------36 7.3 脈衝式電泳結果分析----------------36 7.3.1 圖譜分析------------------36 7.3.2 差異指數(discriminatory index)計算-------37 第四章 結果---------------------38 第一節 菌株鑑定----------------38 1.1 生化鑑定 (API 20NE)---------------38 1.2 聚合酶鏈鎖反應------------38 第二節 抗菌劑感受性試-------------38 第三節 O. rhinotracheale DNA旋轉酶 (DNA gyrase- gyrA gene)之偵測-----------------39 第四節 O. rhinotracheale拓撲異構酶 (Toposiomerase IV parC gene)之偵測-------------------40 第五節 高解析熱熔解分析-----------------40 第六節 脈衝式電泳之分子分型--------------40 第七節 統計與分析----------------41 7.1 菌株抗菌劑感受性百分率統計分析-------------41 7.2 高解析熱溶解分析------------------42 7.3 脈衝式電泳結果分析------------------43 7.3.1 圖譜分析-------------43 7.3.2 差異指數(discriminatory index)------------43 第五章 討論---------------------44 第一節 O. rhinotracheale之抗藥性比較--------------44 第二節 O. rhinotracheale之奎諾酮類抗藥性基因分析-----45 第三節 O. rhinotracheale之分子流行病學--------------47 第六章 結論---------------------51 參考文獻------------------------71 | |
| dc.language.iso | zh-TW | |
| dc.title | 台灣地區禽類鳥鼻氣管桿菌菌株之奎諾酮類抗藥性與gyrA基因之分析 | zh_TW |
| dc.title | Quinolone Resistant Mutations of gyrA Gene of Ornithobacterium rhinotracheale in Poultry in Taiwan | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張紹光(Shao-Kuang Chang),周崇熙(Chung-Hsi Chou),潘銘正(Ming-Cheng Pan),陳秋麟(Chiou-Lin Chen) | |
| dc.subject.keyword | 鳥鼻氣管桿菌,奎諾酮類抗菌劑,gyrA基因,高解析熱熔解分析,脈衝式電泳, | zh_TW |
| dc.subject.keyword | Ornithobacterium rhinotracheale,quinolone antimicrobial,gyrA gene,HRM,PFGE, | en |
| dc.relation.page | 92 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2010-08-10 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
| 顯示於系所單位： | 獸醫學系 | |
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