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研究生:劉昌邦
研究生(外文):Chang-Pan Liu
論文名稱:綠膿桿菌抗藥性基因及抗藥性鮑氏不動桿菌感染的死亡危險因子之分析
論文名稱(外文):THE ANALYSIS OF RESISTANCE GENES OF PSEUDOMONAS AERUGINOSA AND MORTALITY RISK FACTORS OF MULTIRESISTANT ACINETOBACTER BAUMANNII
指導教授:顏聰榮顏聰榮引用關係
指導教授(外文):Tsong-Rong Yan
口試委員:顏聰榮
口試委員(外文):Tsong-Rong Yan
口試日期:2015-01-26
學位類別:博士
校院名稱:大同大學
系所名稱:生物工程學系(所)
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:英文
論文頁數:119
中文關鍵詞:鮑氏不動桿菌綠膿桿菌
外文關鍵詞:A. baumanniiP. aeruginosa
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2013年保加利亞首度發表綠膿桿菌含有超廣效乙內醯胺酶( ESBL)基因blaOXA-142,而後在其它歐洲國家陸續被報導,但在亞洲卻未曾被發表過。本研究則是首度報導台灣綠膿桿菌菌株偵測出超廣效乙內醯胺酶基因blaOXA-142。
從2009年1月至2012年12月,90株綠膿桿菌對ceftazidime(最低抑菌濃度 ≥ 8 mg / L)呈敏感性降低或抗藥性的菌株。以PFGE決定流行菌株的相關聯性,以PCR及基因序列分析的方法來確定超廣效乙內醯胺酶基因的存在。以南方墨點法分析blaOXA-142質體的位置,並執行blaOXA-142的接合和轉形作用。
共有三株綠膿桿菌含有blaOXA-142基因,能產生ESBLs,並呈現ceftazidime抗藥性。MLST顯示它們屬於基因序列型(ST)235,此三株含有blaOXA-142基因的菌株則具有相近的PFGE型別。在第一型挿入子結構中可以偵測出ESBL OXA-142基因。以南方墨點法分析,發現三株blaOXA-142菌株的挿入子是位於質體上,基因分析顯示三株含有blaOXA-142的菌株皆有相同的基因盒。本研究證實在台灣存在blaOXA-142-陽性的綠膿桿菌菌株,三株含有blaOXA-142菌株的基因環境是座落在質體的第一型挿入子。
另一方面,鑑識造成抗碳青黴烯類抗生素之鮑氏不動桿菌感染之死亡風險因子,以及早給予合適的藥物治療,對於病人的預後是很重要的。本研究的目的亦探討182名於台灣醫學中心住院因抗碳青黴烯類抗生素之鮑氏不動桿菌菌血症的患者中,相關死亡風險因子。針對有高風險抗碳青黴烯類抗生素之鮑氏不動桿菌感染及高死亡率之患者,及早投與針對抗碳青黴烯類抗生素之鮑氏不動桿菌有效之抗生素藥物或許可以改善臨床的預後。
The emergence of extended-spectrum β-lactamase (ESBL) OXA-142 gene (blaOXA-142) in Pseudomonas aeruginosa has been reported in Bulgaria and other European countries, but rare in Asia.
From January 2009 to December 2012, the 90 P. aeruginosa isolates with reduced susceptibility or resistance to ceftazidime (MICs ≥ 8 mg/L) were screened for ESBL and other broad-spectrum β-lactamase genes by polymerase chain reaction (PCR) and sequencing. Clonal relationship of the isolates was determined by pulse-field gel electrophoresis (PFGE).
Three isolates were positive for ESBL production, exhibited resistance to ceftazidime and carried the blaOXA-142 gene. The blaOXA-142 gene was integrated into class 1 integron. Using Southern blot analysis, blaOXA-142-containing integron was found to be located in plasmid in all three isolates. Eleven strains of P. aeruginosa carrying blaOXA-17 gene were found. The oprD mutation was found in all the 21 ESBL strains of P. aeruginosa. This study confirmed the presence of blaOXA-142-positive P. aeruginosa isolates in Taiwan.
On the other hand, identification of risks of mortality for carbapenem-resistant Acinetobacter baumannii (CRAB), with early implementation of an appropriate therapy is crucial for the patients' outcome.
The aim of this study was also to survey mortality risk factors in182 patients with CRAB bacteremia in Taiwan. Identification of risks of mortality for CRAB, with early implementation of an appropriate therapy is crucial for the patients' outcome.
ACKNOWLEDGMENTS i
摘要iii
ABSTRACT v
TABLE OF CONTENTS vii
LIST OF TABLES xi
LIST OF FIGURES xii
CHAPTER 1: INTRODUCTION 1
1.1 Research background 1
1.2 Research motive and purposes 9
REFERENCE 10
CHAPTER 2: PSEUDOMONAS AERUGINOSA 12
2.1 Introduction 12
2.1.1 Research background 12
2.1.2 Research motive and purposes 13
2.2 Article review 14
2.3 Material and method 15
2.3.1 Flow chart 15
2.3.2 Bacterial isolates collection, antimicrobial susceptibility testing and patients characteristics 16
2.3.3 Carba NP test 17
2.3.4 Polymerase chain reaction (PCR) and sequencing 19
2.3.5 Pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) 21
2.3.6 Plasmid localization of the blaOXA-142 gene 28
2.4 Results 31
2.4.1 Molecular typing of blaOXA positive P. aeruginosa 31
2.4.2 Susceptibility testing of P. aeruginosa isolates carrying blaOXA-142 35
2.4.3 Characteristics of patients infected with P. aeruginosa isolates carrying blaOXA 35
2.4.4 Detection of oprD mutation 39
2.4.5 Discussion 40
2.4.6 Study limitation 42
2.5 Conclusion and suggestion 43
2.5.1 Conclusion 43
2.5.2 Future suggestion 43
REFERENCE 44
CHAPTER 3: ACINETOBACTER BAUMANNII 50
3.1 Introduction 50
3.1.1 Research background 50
3.1.2 Research motive and purposes 51
3.2 Article review 52
3.3 Material and method 53
3.3.1 Flow chart 53
3.3.2 Bacterial isolates, identification, and clonality determination 54
3.3.3. Study population and data collection 57
3.3.4 Statistical analysis 58
3.4 Results 59
3.4.1 Result 59
3.4.2 Discussion 69
3.4.3 Study limitation 71
3.5 Conclusion and suggestion 72
3.5.1 Conclusion 72
3.5.2 Future suggestion 72
REFERENCE 73
Chinese section 73
English section 73
CHAPTER 4: CONCLUSION 80
APPENDIX 82
Appendix 1: Accepted Article 82
Appendix 2: Article in Press (JMII569 proof Nov. 2014) 83
Appendix 3: Article in Press (JMII proof July 2014) 90
Appendix 4: VITA 97
Appendix 5: Publications List 98
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