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研究生:李永盛
研究生(外文):Yeong-Sheng Lee
論文名稱:痢疾桿菌與百日咳菌分子流行病學
論文名稱(外文):Molecular epidemiology of Shigella flexneri and Bordetella pertussis isolated in Taiwan
指導教授:曾義雄曾義雄引用關係
指導教授(外文):Yi-Hsiung Tseng
學位類別:博士
校院名稱:國立中興大學
系所名稱:分子生物學研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:73
中文關鍵詞:痢疾桿菌百日咳菌分子流行病學脈衝場電泳長期精神照護機構
外文關鍵詞:Shigella flexneriBordetella pertussisMolecular epidemiologyPulsed-field gel electrophoresisLong-Stay Psychiatric Nursing Cente
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本研究係針對台灣常見的桿菌性痢疾及百日咳等兩項傳染病病原菌進行分子流行病學分析。
關於痢疾桿菌部分,本研究收集台灣東部某大型長期精神照護機構持續3年的桿菌性痢疾案件及自社區病人分離的痢疾桿菌進行分子流行病學分析。經利用脈衝場電泳(PFGE, pulsed-field gel electrophoresis)、質體型別(plasmid profile analysis)、抗藥性譜型(antibiogram typing)等方法分析,結果發現該照護機構痢疾桿菌菌株分子次型(molecular subtype)與社區分離株有所不同,且在其相互分隔之數個病房也有其不同分子次型各自衍生並伺機感染致病,而這些菌株可能潛藏在未被偵測到的帶菌者或慢性病人身上。
關於百日咳菌部分,在收集1992~1997年份共114株百日咳菌臨床分離株,進行PFGE分子流行病學分析後,發現1997年台灣出現的百日咳病例流行高峰,是分別由17種PFGE次型所引起,因此排除了台灣出現高傳染性強毒株或抗疫苗株而造成大流行的疑慮。同時因大量病例(71%)接續出現並集中在人口密集的台北都會區,推測擁擠的都市人口有助於疾病的傳播及病原體的演化。

(一) In Taiwan, the number of pertussis cases has been increasing in the recent years, especially in 1997. In addition to the sporadic cases, pertussis also occurred in outbreaks of household, elementary school, and nosocomial infections. Since 71% of reported cases concentrated in the Taipei metropolitan area, it elicited a great deal of concern that a highly contagious strain of Bordetella pertussis might have appeared in Taipei. Therefore, a total of 114 strains of B. pertussis were collected and typed by the molecular method of pulsed-field gel electrophoresis (PFGE). These examined strains include 61 clinical isolates in 1997 and 53 control strains isolated in the previous years (1992~1996). All these strains were classified into 21 subtypes and among them the 1997 collection showed 17 subtypes with 9 new subtypes as compared to those collected in the previous years. This result indicated that the pertussis outbreaks in 1997 were not results of a sole specific virulent strain. It also implies that highly dense population may facilitate the spread and accelerate the genetic divergency of this pathogen.
(二) With 6 separate wards accommodating over 1,600 patients, VNC is a long-stay psychiatric nursing center in eastern Taiwan. During 2001-2003, 39 shigellosis cases occurred in VNC. Different from the notion that most of shigellosis cases are caused by Shigella sonnei, these strains were all S. flexneri except one S. sonnei isolate. O-antigen serotyping showed that the 38 S. flexneri strains were either type 1a (n=20) or 4a (n=18), the two less prevalent serotypes in Taiwan. NotI-PFGE analyses, using 8 1a and 9 4a non-VNC strains isolated during 1996-2003 for comparison, divided the 28 1a and 27 4a strains into 7 and 10 subtypes, designated P1A to P1G and P4A to P4J, respectively. P1A and P4A, appearing in three consecutive years in VNC as well as outside of VNC, are the most prevalent subtypes. Relatedness analyses of the VNC strains based on banding patterns grouped the 1a and 4a strains into 4 and 5 clusters, respectively. The 1a strains, except one, share 95% similarity indicating a common parent, whereas the 4a strains share similarities ranging from 77 to 93% suggesting diverse origins. In two of the outbreaks, less related subtypes of 4a strains were found in the same VNC wards in consecutive years, suggesting possible existence of different subtypes in VNC all the time. Antibiotic susceptibility test showed that the S. flexneri strains are sensitive to at least 7 antibiotics, except one isolate which is sensitive to 3 antibiotics. The data of the latter tests should be helpful for treating S. flexneri infections in Taiwan.

目錄 (Contents) 頁數
目錄 (Contents) 2
摘要 (Abstract) 5
緒言 (Introduction) 7
一. 桿菌性痢疾分子流行病學 9
(一)、摘要 (Abstract) 10
英文 10
中文 12
(二)、前言 (Introduction) 14
(三)、材料與方法 (Materials and Methods) 17
1. 菌種收集 17
2. PFGE操作及親源樹狀圖分析 17
3. 質體型別分析 18
4. 抗藥性譜型分析 19
5. 流行病學資料 19
(四)、結果與討論(Results and Discussion) 20
(五)、結論(Conclusion) 33
(六)、誌謝 (Acknowledgement) 34
(七)、參考資料 (Reference List) 35
(八)、圖表 (Tables and Figures) 41
圖一、病例台灣地理分佈圖 41
圖二、S. flexneri 1a NotI-PFGE親源樹狀分析圖 42
圖三、S. flexneri 4a NotI-PFGE親源樹狀分析圖 43
圖四、質體型別分析圖 44
表一、VNC 2001~2003年病例數及PFGE統計表 45
表二、所有菌株基本資料及分析結果 46
表三、抗藥性譜型結果 48
二. 百日咳菌分子流行病學 49
(一)、摘要 (Abstract) 50
英文 50
中文 51
(二)、前言 (Introduction) 52
(三)、材料與方法 (Materials and Methods) 55
1. 細菌菌株及培養 55
2. PFGE操作及親源樹狀圖分析 55
3. 活體攻擊試驗 56
4. 毒力因子測試 56
5. 南方墨點雜交 56
(四)、結果與討論 (Results and Discussion) 58
(五)、結論 (Conclusion) 62
(六)、誌謝 (Acknowledgement) 63
(七)、參考資料 (Reference List) 64
(八)、圖表 (Tables and Figures) 70
表一.1992~1997年台灣百日咳菌株數及次型分佈 71
圖一.各型百日咳PFGE次型在台灣分佈圖 70
圖二.1992-1994年菌株XbaI-PFGE親源樹狀分析圖 71
圖三.南方墨點雜交結果 72
圖四.1992-2001年菌株XbaI-PFGE親源樹狀分析圖 73

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