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研究生:劉美芳
研究生(外文):Meei-Fang Liu
論文名稱:台灣地區抗甲氧苯青黴素金黃色葡萄球菌的流行病學分析
論文名稱(外文):Epidemiology of methecillin resistant Staphylococcus aureus isolates in Taiwan
指導教授:林赫
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:2008
畢業學年度:96
語文別:中文
論文頁數:63
中文關鍵詞:抗甲氧苯青黴素金黃色葡萄球菌脈衝電場電泳基因分型法多基因組序列分析院內感染族群
外文關鍵詞:MRSAPFGEMLSTclusternosocomial infection
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摘 要
金黃色葡萄球菌(Staphylococcus aureus)是醫院最常見的致病菌,自從1961年分離出第一株抗甲氧苯青黴素金黃色葡萄球菌(methecillin resistant Staphylococcus aureus;MRSA)之後,MRSA很快的出現在各國,由於抗生素的大量使用,使MRSA成為目前院內感染最常見的菌種。本研究收集台灣地區5家醫院自2001年1月至2008年3月共107株MRSA,期望藉脈衝電場電泳基因分型法(Pulsed-field gel electrophoresis;PFGE)、多基因組序列分析(Multilocuse sequence typing;MLST)兩種方法,分析菌株的基因相關性,並進一步和其他國家比較,找出台灣的本土流行株,並比較台灣不同地域MRSA的親源相關性。脈衝電場電泳結果,經比對分析菌株親緣樹枝圖後,親緣百分比>80%即定義為同一族群(cluster),因此得到五種族群:A(80.5%)、B(89.8%)、C(80.4%)、D(88.0%)與E(87.5%),其中A群與B群全台5家醫院都存在此兩群,C、D、E群則各分布在不同醫院。本次107株MRSA經MLST分型再與資料庫比對,共87株可比對,且有6種型別,包括:ST239、ST59、ST241、ST5、ST25與ST221,最主要的是ST239與ST59,各佔59.8%與13%。從本研究發現ST239是臺灣最主要的流行株,與亞洲國家有些異同,如韓國主要的流行株是ST5與ST239,而日本主要的流行株是ST5,又中國的主要菌株是ST239,和台灣具相同的流行株,原因可能是中國和台灣經常有台商和探親來往頻繁之故。另一方面,本研究呈現MRSA在臺灣的醫院是廣泛分佈的,所以醫院對於此抗藥性菌株,除了持續的監測,完備的感控措施及完善的抗生素管制外,嚴謹的院內感染管制政策的落實,是最重要的預防院內感染(nosocomial infection)之道。
目 次
誌謝--------------------------------------------------------------- i
中文摘要----------------------------------------------------------- ii
英文摘要-------------------------------------------------------- iii
目次------------------------------------------------------------------------- iv
圖表次----------------------------------------------------------------------- v
壹、前言-------------------------------------------------------------------- 1
貳、材 料 與 方 法
一、菌株來源-------------------------------------------------------------- 6
二、藥品-------------------------------------------------------------------- 7
三、培養基----------------------------------------------------------------- 7
四、抗生素----------------------------------------------------------------- 8
五、細菌之培養與鑑定-------------------------------------------------- 8
六、最低抑菌濃度-------------------------------------------------------- 11
七、脈衝電場電泳法----------------------------------------------------- 12
八、PFGE條紋片段的比對---------------------------------------------- 14
九、DNA萃取--------------------------------------------------------------- 15
十、進行MLST前的PCR反應------------------------------------------- 15
十一、多基因定序分型-------------------------------------------------- 16
十二、mecA測定---------------------------------------------------------- 17
叁、結果
一、細菌鑑定-------------------------------------------------------------- 18
二、最低抑菌濃度-------------------------------------------------------- 18
三、PCR反應--------------------------------------------------------------- 19
四、脈衝電場電泳-------------------------------------------------------- 19
五、多基因定序分型----------------------------------------------------- 20
肆、討論-------------------------------------------------------------------- 21
伍、結論-------------------------------------------------------------------- 27
陸、參考文獻-------------------------------------------------------------- 48
柒、附錄-------------------------------------------------------------------------------- 63





















圖 表 次
表一金黃色葡萄球菌對oxacillin的最低抑菌濃度結果-------- 30
表二、MLST PCR引子的序列------------------------------------------ 31
表三、MLST基因型的表現--------------------------------------- 32
表四、台灣地區醫院ST的分布--------------------------------------- 33
表五、臺灣地區PFGE型別的分佈------------------------------------ 34
圖一、金黃色葡萄球菌在血液培養基的菌落型態----------------- 35
圖二、革蘭氏染色結果-------------------------------------------------- 36
圖三、tpi、gmk與pta為引子的PCR產物--------------------------- 37
圖四、mecA為引子的PCR產物--------------------------------------- 38
圖五、MRSA的脈衝電場電泳結果------------------------------------- 39
圖六、107株MRSA的PFGE結果親緣樹枝圖------------------------ 40
圖七、中榮MRSA的PFGE結果親緣樹枝圖------------------------ 41
圖八、馬偕MRSA的PFGE結果親緣樹枝圖-------------------------- 42
圖九、桃園MRSA的PFGE結果親緣樹枝--------------------------- 43
圖十、屏東MRSA的PFGE結果親緣樹枝圖------------------------ 44
圖十一、高雄MRSA的PFGE結果親緣樹枝圖-------------------- 45
圖十二、ST59的PFGE結果親緣樹枝圖------------------------------ 46
圖十三、ST239的PFGE結果親緣樹枝圖---------------------------- 47
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