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研究生:陳昶華
研究生(外文):Chang-Hua Chen
論文名稱:包氏不動桿菌的分子特性與流行病學之研究
論文名稱(外文):Molecular Characterization and Epidemiological Study of Clinical Acinetobacter baumannii
指導教授:黃介辰林清淵林清淵引用關係
指導教授(外文):Chieh-Chen HuangChing-Yuang Lin
學位類別:碩士
校院名稱:國立中興大學
系所名稱:生命科學院碩士在職專班
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:170
中文關鍵詞:包氏不動桿菌IntI1bla Tem—1表現型基因型
外文關鍵詞:Acinetobacter baumanniiIntI1 genebla Tem—1 genegenotypephenotype
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近年彰化基督教醫院出現具有imipenem抗藥性的包氏不動桿菌(Acinetobacter baumannii) 後,包氏不動桿菌就被引起注意。加上抗生素抗藥性的問題日益嚴重,因此本研究主要目的首要建立彰化基督教醫院等中部地區包氏不動桿菌的分子流性病學資料庫,其次分析表現型與基因型的相關性,及探討抗藥性基因(bla Tem—1與 IntI1)的水平傳播功能。以民國90年四月1日到民國91年三月31日為止,在彰化基督教醫院分離出之401 株包氏不動桿菌為材料,以PCR方法進行抗藥性基因( bla Tem—1與 IntI1基因)之研究。依照finite mixture方法,將菌株的抗生素抗藥性表現型分為三組(clusters)32群(subgroup)。帶有bla Tem-1 基因者佔31.2 % (10/32) ,與帶有IntI1 基因者佔81.3% (26/32)。其次吾人也發現E. coli (AF427133)與 A. baumannii CH 7的bla Tem-1 基因有100%相似性(1044/1044)。根據彰基對此菌株的SXT 藥物敏感實驗結果,發現81.3% (26/32)帶有IntI1 基因,其中 84.6%( 22/26) IntI1 陽性菌株中會對SXT有抗藥性,也發現其中80.8% (21/26)IntI1 陽性菌株中會對gentamicin有抗藥性。根據彰基對此菌株的CAZ 藥物敏感實驗結果,發現31.2% (10/32)帶有bla Tem-1 基因,其中 100%( 10/10) bla Tem-1陽性菌株會對CAZ有抗藥性。本研究的結論為包氏不動桿菌的IntI1 基因可能跟 SXT 抗藥性有關係,與bla Tem-1基因可能跟 CAZ 抗藥性有關係。關於遺傳情報水平傳播的部分有待進一步的研究。

The Acinetobacter baumannii was noticed since imipenem-resistant had emerged recently at Changhua Christian Hospital, Changhua, Taiwan. Because of the growing importance of antibiotic resistant strains of A. baumannii, this study firstly established the native clinical epidemiological database of A. baumannii strains in the central Taiwan area, secondarily analyzed the relationship between genotype (bla Tem—1, and IntI1 gene) and the antibiotic resistance phenotype, and studied the horizontal gene transfer of the bla Tem—1. A cross-sectional, prospective study was conducted at Changhua Christian Hospital during April 1, 2001, and March 31, 2002. A total of 401 clinical A. baumannii strains were collected. The PCR method was used to detect the target resistant genes (bla Tem-1 gene , IntI1 gene). We divided the 401 clinical strains into the 3 clusters and 32 subgroups according to the antibiotics resistant patterns by the finite mixture method. 31.2 % (10/32) A. baumannii possessed the bla Tem-1 gene, while 81.3% (26/32) possessed the IntI1 genes. The DNA sequence of bla tem gene was identical (1044/1044 bp) between the E. coli (AF427133) and the A. baumannii CH 7. According to the SXT susceptibility tests and PCR analysis of the Changhua Christian Hospital, 81.3% (26/32) of clinical strains carried the IntI1 gene, and 84.6%( 22/26) IntI1 positive strains were resistant to SXT. 80.8% (21/26) in IntI1 positive strains were resistant to gentamicin and 16.7%(1/6) in IntI1 negative strains were resistant to gentamicin. The IntI1 gene could result in the SXT and gentamicin resistance. Furthermore, CAZ susceptibility tests and PCR analysis showed that 31.2% (10/32) of clinical strains carried the bla Tem-1 gene, and 100%( 10/10) bla Tem-1 positive strains were CAZ resistant. We concluded that the IntI1 genes could relate to the SXT resistance, and the bla Tem—1 gene could relate to the CAZ resistance. The horizontal gene transfer of the bla Tem—1 gene needed further experiments

Preface
Abstracts
Chapter One: Introduction (1)
Chapter Two: Molecular Epidemiological Study of Clinical Acinetobacter baumannii Isolates from Changhua Christian Hospital
Background (9)
Purpose of study (11)
Materials and methods (11)
A) Strategy
B) Materials
1) Bacterial strains
2) Clinical data collection
C) Methods
1) Microbiological identifications
2) Susceptibility tests
3) Protocols of DNA extraction
4) Choice of the target resistant genes
5) Design of the primers and PCR conditions
a) Molecular detection of IntI1 gene
b) Molecular detection of bla Tem-1 gene
6) DNA sequencing
7) Alignment
8) Statistical methods
Results (17)
A)Antibiotics resistant phenotype and grouping
B) Antibiotics resistant genotype
C) DNA sequence and alignment
Discussions (24)
Chapter Three: Trimethoprim-sulfamethoxazole and Aminoglycosides Resistance Analysis of the IntI1 gene of Clinical Acinetobacter baumannii Isolates from Changhua Christian Hospital
Background (30)
Purpose of study (33)
Materials and Methods (33)
A) Strategy
B) Materials and methods
1) Bacterial strains
2) Susceptibility tests
3) Characterization of IntI1 gene of Clinical Acinetobacter baumannii Isolates
4) Statistical Analysis
Results (34)
Discussion (35)
Chapter Four: Ceftazidime Resistance of the bla Tem-1 gene of Clinical Acinetobacter baumannii Isolates from Changhua Christian Hospital
Background (38)
Purpose of study (40)
Materials and Methods (40)
A) Strategy
B) Materials and methods
1) Bacterial strains
2) Susceptibility tests
3) The characterization of the bla Tem-1 gene of Clinical Acinetobacter baumannii
4) Cloning
5) Statistical Analysis
Results (42)
Discussion (44)
Chapter Five: Conclusions (47)
The ongoing study (48)
References (49)
Tables
Figures
Appendix
Abbreviations
Acknowledge
中文前言
中文摘要
第一章 簡介 (1)
第二章 彰化基督教醫院所分離之包氏不動桿菌的分子流行病學研究 (9)
研究目的 (11)
材料與方法 (11)
A) 研究策略
B) 材料
1) 菌株來源
2) 臨床資料
C) 方法
1) 微生物學鑑定方法
2) 抗生素藥物敏感性實驗
3) 染色體抽取方法
4) 目標基因的選擇
5) PCR 的條件
a) IntI1 基因的偵測
b) bla Tem-1基因的偵測
6) DNA 定序
7) DNA比對
8) 統計方法
結果 (17)
A) 抗生素抗藥性表現型
B) 抗生素抗藥性基因型
C) DNA比對
討論 (24)
第三章 彰化基督教醫院包氏不動桿菌的trimethoprim-sulfamethoxazole 與 aminoglycosides 抗藥性與IntI1基因的特性分析
背景 (30)
研究目的 (33)
材料與方法 (33)
A) 研究策略
B) 材料與方法
1) 微生物學鑑定方法
2) 抗生素藥物敏感性實驗
3) IntI1 基因的特性分析
4) 統計方法
結果 (34)
討論 (35)
第四章彰化基督教醫院包氏不動桿菌的ceftazidime 抗藥性與bla Tem-1基因的特性分析
背景 (38)
研究目的 (40)
材料與方法 (40)
A) 研究策略
B) 材料與方法
1) 微生物學鑑定方法
2) 抗生素藥物敏感性實驗
3) bla Tem-1基因的特性分析
4) Cloning
5) 統計方法
結果 (42)
討論 (44)
第五章 結論 (47)
未來的計畫 (48)
參考文獻 (49)
表格
圖片
附錄
縮寫
中文致謝

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