Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/22376
標題: Class 1 integron在鮑氏不動桿菌臨床分離株中的盛行率與其水平傳播單位的解析
Prevalence of class 1 integron among clinical isolates of Acinetobacter baumannii and molecular characterization of its horizontal mobile unit.
作者: 朱韋臣
Chu, Wei-Chen
關鍵字: Acinetobacter baumannii
鮑式不動桿菌
antibiotic resistance
integron
resistande island
抗生素抗性
插入子
抗性基因島嶼
出版社: 生命科學系所
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摘要: 近年來帶有多重抗藥性的鮑式不動桿菌逐漸成為各國醫院內感染的主要病原菌之ㄧ。而integron是近年來被認為除了質體和轉位子外,另一個與細菌獲得新抗藥基因有關的水平傳播單位。本篇研究針對最常見的class 1 integron,其5’及3’保留性序列設計的引子,針對彰化基督教醫院的32株鮑式不動桿菌,以及大林慈濟醫院的18株全抗藥性的鮑式不動桿菌,進行PCR的檢測,在彰化基督教醫院的菌株中有65.6%(21/32)的菌株發現有class 1 integron的存在,並帶有四種大小不同的cassette array片段。分別是3.0 kb (aacC1-orfX- orfX-orfX´- aadA1a)、2.5 kb(aacA4-catB8-aadA1)、2.0kb(dhfrXII-orfF- aadA2)、1.2 kb (dhfrAI- ORF)。而在大林慈濟醫院的全抗性菌株中100%發現到有class 1 integron,只有三種不同的大小,分別是3.0 kb、2.5 kb以及2.0 kb。對照抗生素的抗性測試可以發現,帶有Class 1 integron的菌株,其具有的抗生素抗性比不具有Class 1 integron的菌株廣泛,但是這些integron所挾帶的cassette array上的抗性基因種類卻只有其中一部分。而發現頻率最高的為2.5 kb的integron。為2004年在彰化基督教醫院鮑式不動桿菌菌株CH7上所發現,當時為世界首見,並且發現台灣醫院內分離的鮑式不動桿菌菌株帶有此integron的情況很普遍,但是其他國家的發現此integron的報告卻極少。為了了解該integron的特性及所在位置,利用Inverse PCR的方式進行針對CH7 2.5 kb integron 的上下游DNA序列解析,發現其上下游都帶有aphA1(kanamycin抗性基因),以及transposase gene tnpIS15/ tnpAIS26,發現這些序列和法國鮑式不動桿菌菌株AYE的 86 kb 的resistance island(目前報導過的最大抗性基因群)的部分序列相當類似,所以推測帶有2.5 kb cassette array的菌株可能帶有類似resistance island的結構。為了檢測其他菌株是否有類似的情況,針對其他帶有2.5 kb integron的菌株進行PCR- RFLP的分析,發現上游序列(aphA1 -tnpIS15/ tnpAIS26)和下游序列(orf5-tnpIS15/ tnpAIS26)完全相同者,也同時帶有3.0 kb的cassette array。縱合以上資料推測2.5 kb及3.0 kb cassette若同時出現可能位於同一個resistance island結構,而只帶有2.5 kb cassette array的菌株,其結構不相同。帶有2.5 kb integron的鮑式不動桿菌,可能經由長期抗生素壓力的篩選,形成台灣地區特有的臨床優勢菌株。
The multi-drug resistant Acinetobacter baumannii had became one of the major infectious pathogens in hospital around the world. Since integrons are the genetic unit that confer antibiotic resistance to bacteria besides plasmid and transposon, our study focused on the class 1 integron that was the most commonly found in bacteria. Bacterial strains were collected from Changhua Christian Hospital and Dalin buddist Tzu Chi Hospital. The CS-PCR method was used to detect the class 1 integrons , cassette array sizes and their sequences in these strains. It was found that 65.5 % of the Changhua Christian Hospital strain carried the class 1 integron, and showed only 4 kinds of different size cassette arrays: 3.0 kb (aacC1-ORFX- ORFX-ORFX´- aadA1a), 2.5 kb (aacA4-catB8- aadA1), 2.0kb(dhfrXII -orfF-aadA2) and 1.2 kb (dhfrAI- ORF). There were 100% of the Dalin buddist Tzu Chi Hospital strain carried the class 1 integron, and only 3 kinds of different size cassette arrays: 3.0 kb, 2.5 kb and 2.0kb. Compared with the antibiotic sensivity test data, we found that multi drug resistance usually carried within integron, but the resistance gene on the cassette array were only a small part of the resistance phenotype . The most common integron found in this study were the size of 2.5 kb cassette arrays. It was initially found in the A. baumannii CH7 at Changhua Christian Hospital.It was very common in A. baumannii isolated from hospital in Taiwan, but rarely reported in other country. In order to understand the 2.5 kb integron's characterization, we use the Inverse PCR method to analysis the sequence that upstream and downstream of the 2.5 kb integron in CH7. We found that upstream or downstream both carried the aphA1 (kanamycin resistance gene) and transposase gene tnpIS15/tnpIS26, and the blast program analysis show that these sequences demonstrate high similarity with the resistance island in the A. baumannii AYE from French. By using the PCR-RFLP method to check the other strains that carried 2.5 kb integron, we found that the strains carried 2.5 kb and 3.0 kb integron were identical with CH7. The data reported here supports that the strain have 2.5 kb integron may pass through the antibiotic pressure selection and became a dominant strain in Taiwanese Hospitals.
URI: http://hdl.handle.net/11455/22376
其他識別: U0005-2207200601515500
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2207200601515500
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