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標題: 使用脈衝式電泳分析及插入子聚合酶鏈鎖反應對多重抗藥性鮑氏不動桿菌進行分子流行病學的分析
Molecular Analysis of Epidemiology of Multiple Drug Resistance Acinetobacter baumannii: by Pulse-Field Gel Electrophoresis, and Integron Polymerase Chain Reaction.
作者: 陳玉娟
Chen, Yu-Chuan
關鍵字: Multiple Drug Resistance Acinetobacter baumannii;多重抗藥性鮑氏不動桿菌
出版社: 生命科學院碩士在職專班
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不動桿菌屬(Acinetobacter spp.)是革蘭氏陰性球桿菌,為常見的院內感染菌它含有17個基因種(genospecies) 。此菌廣範的分佈在自然界中,包括醫院的環境,對於長期住院病人它可變成皮膚及呼吸道菌叢的ㄧ部份。此菌傳播的方式最常見的原因為環境因素造成住院病人的移生菌colonizaltion) ,然後藉由醫療器具如﹕血管內管或導尿管,導引微生物至正常無菌的部位。
在臨床上分離的鮑氏不動桿菌(Acinetobacter baumannii)常為移生菌,真正的感染為院內感染,最常見的包括生殖泌尿道(genitourinary tract) 、呼吸道(respiratory tract) 、傷口(wound) 、軟組織(soft tissues )及菌血症(bacteremia) 。鲍氏不動桿菌(Acinetobacter baumannii)在免疫機能低下的病人如肺炎、尿道炎、心內膜炎、傷口感染、腦膜炎與敗血病….等,是很常見的院內感染病原菌。在Carbapenems類藥物中主要的Imipenem and Meropenem,是治療因感染鲍氏不動桿菌有效的治劑。 本研究以嘉義縣大林慈基醫院及鄰近醫院收集的MDRAB 臨床菌株之流行菌株為基礎,分析其抗生素的表現型與基因型的相關性,並探討其抗藥基因是否經由水平傳播的方式散布於各菌株間之可能性。使用的臨床菌株是在民國九十三年一月一日到民國九十四年十二月三十一日止,在大林慈濟醫院及鄰近醫院收集的19株鮑氏不動桿菌(Acinetobacter baumannii)包括15株MDRAB 4株non-MDRAB為材料,分別以Disck diffusion experiment 、E- test與BD PhoenixTM 等抗生素藥物敏感性試驗來測定這19株菌株的抗生素抗藥性表現型;另外以脈衝電場電泳(PFGE) 電泳後用圖普分析儀軟體-Bionumerics進行比對描繪菌株親緣關係。PFGE結果顯示這19株菌株分成17種類型而鑑別力達0.99%。分析15株MDRAB中N1、N2、N3、N5、N6、N7、N8、N9、N10、N12、N17、N18、N19間的相似性>80%,而N17、N18、N19是鄰近醫院分離出的MDRAB菌種,實驗結果證實MDRAB菌種在院際間傳播的可能性。 另外以聚合酶連鎖反應(PCR) 方法進行Cabapenem 的抗藥基因(使用的引子為blaOXA23 F/R、blaOXA24 F/R、blaVIM-1 F/R、blaVIM-2 F/R、blaIMP-1 F/R等五對引子)與水平傳播平台Integron內含的抗藥基因來分析,結果顯示這15株MDRAB中並未發現與Cabapenem 的抗藥基因有關之特異性產物;但這15株MDRAB均帶有均帶有一個約2.5Kb的第一類Integron,其中的cassette排列方式也相同這結果顯示出抗藥基因水平傳播的方式。

Acinetobacter spp. is Gram-Negative bacilli or coccobacilli and there is common nosocomial pathogen, particular among immunocompromised patients. There are 17 genospecies in the genus Acinetobacter. Acinetobacter spp. is widely distributed in nature, including the hospital environment. They may become established as part of skin and respiratory flora of patients. Mode of transmission is colonization of hospitalizated patients from environmental factors; medical instrumentation (e.g., intravenous or urinary catheters) introduces organism to normally sterile sites. Acinetobacter spp. is common environmental opportunistic pathogens, they can be found on the skin and mucosa of healthy people. But they cause lots of serious infectious disease such as pneumonia, urinary tract infection, endocarditis infection, wound infection, meningitis infection and septicemia, especially in immunocompromised patients. Carbapenems, maily imipenem and meropenem, are potent agents nts for the treatment of infections due to multiple resistant Acintobacter baumannii. Based on the epidemic infectiomation on clinical MDRAB strains collected by Buddhist Dalin Tzu Chi General Hospital, Chiayi, Taiwan, and neighborhood hospital correlation between their phenotype and genotype is analyzed in this research. In addition, the possibility of whether the drug-resistant gene diffuses among strains in a horizontal way is strains used in this study were clinical strains isolated from in Buddhist Dalin Tzu Chi General Hospital and neighborhood hospital from January 1, 2004 to December 31, 2005. They were further conducted an antibitc susceptibility test by the Disk diffusion experiment and BD PhoenixTM system to explored. The phenotype of these fifteen imipenem-resistance and four imipenem-susceptible strain. Furthermore the “Bionumerics” analysis software was applied after PFGE to compare and locate the genetic/phylogenesis relationship among the strains. The PFGE result indicated those strain could be divided into 17 types. Besides, drug-resistant genes of Carbapenem(blaOXA-23、blaOXA-24、 blaIMP-1、blaVIM-1、blaVIM-2、 blaTEM-1 ) and those of integron, a horizontal diffusion platform, were analyzed with the PCR method. No specificity PCR products found to PCR products were found to the Carpapenem-resistant related genes of the fifteen isolated strains. However, there was a type 1 integron of about 2.5 Kb existed in all these fifteen MDRAB strains and the arrangement of cassette was the same. The result revealed the possibility of horizontal transfer of the drug-resistant genes.
其他識別: U0005-0508200613101000
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