文章摘要
武 渊,李 慧,梅亚宁,李 军,文 怡,黄祖瑚.在铜绿假单胞菌临床分离株1类整合子中发现携带新型耐药基因盒[J].南京医科大学学报,2008,28(1):37~41
在铜绿假单胞菌临床分离株1类整合子中发现携带新型耐药基因盒
Identification of class 1 integron carrying novel gene cassettes in Pseudomonas aeruginosa isolates
投稿时间:2007-08-22  
DOI:10.7655
中文关键词: 假单胞菌,铜绿  1类整合子  基因盒  耐药
英文关键词: Pseudomonas aeruginosa  integron  gene cassette  resistance
基金项目:江苏省临床医学重点学科基金资助
作者单位
武 渊 南京医科大学第一附属医院感染病科,江苏 南京 210029 
李 慧 南京医科大学第一附属医院感染病科,江苏 南京 210029 
梅亚宁 临床检验中心,江苏 南京 210029 
李 军 南京医科大学第一附属医院感染病科,江苏 南京 210029 
文 怡 临床检验中心,江苏 南京 210029 
黄祖瑚 南京医科大学第一附属医院感染病科,江苏 南京 210029 
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中文摘要:
      目的:研究铜绿假单胞菌中1类整合子的分布和结构特征及与多重耐药的关系?方法:应用聚合酶链反应(PCR)检测铜绿假单胞菌1类整合酶基因(intⅠ)并扩增其可变区;DNA测序技术分析1类整合子可变区的耐药基因;对1类整合子阳性菌采用琼脂平皿稀释法测定其MIC并进行转移接合明确其耐药基因转移情况;脉冲场凝胶电泳法(PFGE)了解这些阳性菌的遗传相关性?结果:47株铜绿假单胞菌临床分离株中,29株携带1类整合子,未检测到3类整合子?这29株包含的整合子均扩增出长度约4.3 kb的可变区,核酸序列分析发现存在新型耐药基因盒组合形式aac(6’)-Ⅱ-aadA13-cmlA8-oxa-10,Genbank基因号为EU182575?其中基因盒aadA13以往未在铜绿假单胞菌中发现?同时第3个开放阅读框与已知基因cmlA5有95%的同源性,为编码氯霉素外排泵的基因盒,将其命名为cmlA8?PFGE结果分析显示29株阳性菌株具有高度的同源性?结论:本院流行的铜绿假单胞菌携带包含新型基因盒及组合形式的Ⅰ类整合子,再次证实1类整合子参与了铜绿假单胞菌的耐药和多重耐药?
英文摘要:
      Objective:To investigate the distribution,characterization of class 1 integron and its action in multi-resistance in Pseudomonas aeruginosa. Methods:PCR and DNA sequencing methods were undertaken to clarify the type and context of variable region of class 1 integron;the susceptibility of class 1 integron positive strains to five represented antibiotics was determined by agar dilution method;conjugation experiment was carried out to disclose its transferable property;and lastly,pulsed-field gel electrophoresis pattern was performed to explain the genetic relationship of those class 1 integron positive Pseudomonas aeruginosa. Results:Of 47 isolates,29 strains of Pseudomonas aeruginosa were found containing class 1 integron,and no class 3 integron was detected. A length of 4.300 bp variable region was found in all the class 1 integron positive strains by PCR amplification. The resulting DNA sequences revealed that it was comprised of a new array of aac(6’)-Ⅱ-aadA13-cmlA8-oxa-10 gene cassettes,and the genbank number is EU182575. According to the database in genbank,aadA13 was previously unreported in Pseudomonas aeruginonsa; chloramphenicol efflux pumping determinant,named as cmlA8 shares 95% nucleotide similarity to cmlA5 gene. All those class 1 integron positive isolates showed high homology by PFGE pattern analysis after digestion with SpeⅠ. Conclusion:Class 1 integrons disseminated among correlated Pseudomonas aeruginosa in our hospital possessing novel gene cassettes and mediated the resistance and multi-drug resistance.
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