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研究生:張家瑋
研究生(外文):Chia-Wei Chang
論文名稱:2011年12月至2013年11月國立臺灣大學生物資源暨農學院附設動物醫院從狗與貓所分離出具廣效性乙內醯胺細菌株的分離率及特性分析
論文名稱(外文):Separation rate and characteristics of extended-spectrum β-lactamase-producing bacterial isolates from dog and cat in National Taiwan University Veterinary Hospital during December 2011 to November 2013
指導教授:蔡向榮蔡向榮引用關係
口試委員:張紹光周崇熙陳德勛
口試日期:2014-06-24
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:獸醫學研究所
學門:獸醫學門
學類:獸醫學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:71
中文關鍵詞:抗生素抗藥性腸內菌科超廣效性乙內醯胺&;#37238;多基因位點序列分析頭孢菌素
外文關鍵詞:antimicrobial resistanceEnterobacteriaceaeextended-spectrum β-lactamasesmultilocus sequence typingcephalosporins
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自從1980年代初期,廣效性乙內醯胺&;#37238;已逐漸成為抗生素抗藥性領域研究的新課題。廣效性乙內醯胺&;#37238;可以抵抗乙內醯胺類抗生素並被乙內醯胺&;#37238;抑制劑抑制。研究顯示,大腸桿菌與肺炎克雷伯氏菌為最常見到的廣效性乙內醯胺&;#37238;產生菌株。近年各國研究均指出帶有廣效性乙內醯胺&;#37238;的菌株分離率正逐漸增加,而且傳播型態也逐漸改變。相對而言,台灣在伴侶動物所分離的菌株是否帶有廣效性乙內醯胺&;#37238;及其特性等相關資訊則較有限。
本研究之目的在於檢查並分析由2011年12月至2013年11月間,由國立臺灣大學生物資源暨農學院附設動物醫院從犬貓所分離到的臨床菌株是否帶有廣效性乙內醯胺&;#37238;及細菌的相關性狀分析。依照美國臨床和實驗室標準學會 (CLSI)之雙紙錠協同測試法確認步驟進行廣效性乙內醯胺&;#37238;表現型偵測。研究結果顯示465株菌株中共有60株細菌偵測到表現廣效性乙內醯胺&;#37238;性狀。這裡面包含了26株大腸桿菌、16株肺炎克雷伯氏菌、10株腸桿菌屬細菌、3株不動桿菌屬細菌、3株沙雷氏菌屬細菌與2株綠膿桿菌。整體菌株之廣效性乙內醯胺&;#37238;分離率為12.9%。TEM和CTX-M是最主要的廣效性乙內醯胺&;#37238;基因型,在CTX-M的次分型中則以CTX-M-1次分群最常見。多基因位點序列分析結果顯示在可產生廣效性乙內醯胺&;#37238;的大腸桿菌中最常分離到的序列型為ST457。
研究結果顯示在國立臺灣大學生物資源暨農學院附設動物醫院的伴侶動物廣效性乙內醯胺&;#37238;的分離率是相對高於西方國家的,抗藥性的增加也暗示著在臨床治療上的困難。基因分型和多基因位點序列分析結果皆顯示在台灣伴侶動物廣效性乙內醯胺&;#37238;與人醫之間的差異。本結果僅代表國立臺灣大學生物資源暨農學院附設動物醫院之伴侶動物中廣效性乙內醯胺&;#37238;流行病學研究結果,無法代表台灣整體伴侶動物的廣效性乙內醯胺&;#37238;流行病學研究結果。


Since the early 1980s, extended-spectrum β-lactamases (ESBLs) had become an emerging issue for antibiotic resistance. Production of ESBL confers resistance to the commonly used beta-lactam antimicrobials, but are inhibited by β-lactamases inhibitors. Escherichia coli and Klebsiella pneumoniae are the two major ESBL-producing bacteria. Several studies have revealed that the ESBL prevalence around the world is increasing in recent years, and the transmission route is changed. On the contrary, the isolation rates of resistance to ESBL among microorganisms from companion animals remained limited in Taiwan.
The objective of the present study was to examine and characterize the occurrence of ESBL-producing microorganisms in clinical samples of dogs/cats attending to National Taiwan University Veterinary Hospital (NTUVH) during December 2011 to November 2013. Combination disc tests using cefotaxime and ceftazidime (30 μg) disks, with and without clavulanic acid (10 μg) according to CLSI guidelines resulted in 60 isolates demonstrating an ESBL phenotype from 465 isolates included in this study. Twenty-six Escherichia coli, 16 Klebsiella pneumoniae, 10 Enterobacter spp., 3 Acinetobacter spp., 3 Serratia spp, and 2 Pseudomonas aeruginosa isolates exhibited ESBL phenotype. The ESBL prevalence was 12.9%. TEM and CTX-M were the two major ESBL genes, while CTX-M-1 was the most commonly found subgroup in CTX-M subtyping. Multilocus sequence typing (MLST) revealed that ST457 was the predominant sequence type in ESBL-producing E. coli.
Our result showed that the isolation rate of ESBL in NTUVH is comparatively higher than that from western countries. Increase of drug resistance may indicate the treatment difficulty in clinical practice. Gene typing and MLST results both showed the difference between human and companion animals in ESBL. Our results only represent the epidemiology of ESBL-producing bacteria from the companion animals in NTUVH, not the whole Taiwan’s companion animals.


中文摘要………………………………………………………………… I
英文摘要…………………………………………………………………II
目錄………………………………………………………………………IV
表目錄…………………………………………………………………VII
圖目錄…………………………………………………………………VIII
第一章 緒論…………………………………………………………1
第二章 文獻探討……………………………………………………3
2.1 腸內菌科細菌簡介…………………………………………………3
2.1.1大腸桿菌…………………………………………………………3
2.1.2肺炎克雷伯氏菌…………………………………………………4
2.1.3沙門氏菌…………………………………………………………5
2.1.4奇異變形桿菌……………………………………………………7
2.2 β-lactam類抗生素…………………………………………………8
2.2.1盤尼西林類………………………………………………………8
2.2.2碳青黴烯類………………………………………………………9
2.2.3頭孢菌素類………………………………………………………9
2.4單環胺基類…………………………………………………………10
2.3細菌之抗藥性機制及傳播…………………………………………10
2.3.1使藥物無法在細菌內累積……………………………………10
2.3.2使藥物作用的位置改變………………………………………11
2.3.3使藥物分解或失去活性………………………………………11
2.4 乙內醯胺&;#37238;…………………………………………………………12
2.4.1 Ambler分類原則…………………………………………………12
2.4.2 Bush分類原則……………………………………………………13
2.5 乙內醯胺&;#37238;抑制劑…………………………………………………13
2.6 ESBL相關基因及偵測方式…………………………………………14
2.6.1 TEM………………………………………………………………15
2.6.2 SHV………………………………………………………………15
2.6.3 CTX-M……………………………………………………………15
2.6.4 表現型偵測………………………………………………………16
2.6.5 基因型偵測………………………………………………………17
2.7 ESBL之流行病學調查結果…………………………………………17
2.7.1 ESBL國際流行病學調查結果…………………………………17
2.7.2 ESBL在台灣的研究……………………………………………19
第三章 研究目的………………………………………………………20
第四章 材料與方法……………………………………………………21
4.1樣本菌株之來源……………………………………………………21
4.2 ESBL鑑定……………………………………………………………21
4.3 ESBL陽性菌株染色體DNA萃取……………………………………22
4.4細菌ESBL抗藥基因分型……………………………………………23
4.5基因定序 (DNA sequencing)………………………………………23
4.6多基因位點序列分析 (Multilocus sequence typing, MLST)……………25
第五章 研究結果………………………………………………………26
5.1 ESBL菌株分離率……………………………………………………26
5.2 ESBL菌株分型………………………………………………………26
5.3 ESBL菌株抗藥性分析………………………………………………27
5.4多基因位點序列分析結果 (MLST)…………………………………28
第六章 討論……………………………………………………………29
6.1分離率調查結果………………………………………………29
6.2 ESBL-producing菌株基因型分析……………………………30
6.3 ESBL-producing菌株抗藥情形………………………………32
6.4多基因位點序列分析…………………………………………34
第七章 結論……………………………………………………………35
參考文獻…………………………………………………………………47


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