同一患者不同部位的4株肺炎克雷伯菌对碳青霉烯类抗生素耐药机制研究和同源性分析

被引:10
作者
刘瑛 [1 ]
俞静 [1 ]
张良 [2 ]
沈立松 [1 ]
机构
[1] 上海交通大学医学院附属新华医院检验科
[2] 上海交通大学医学院附属新华医院泌尿外科
关键词
肺炎克雷伯菌; 碳青霉烯酶; 外膜蛋白;
D O I
10.16718/j.1009-7708.2013.06.009
中图分类号
R446.5 [微生物学检验];
学科分类号
100208 ;
摘要
目的对临床分离自同一患者不同部位的4株耐碳青霉烯类抗生素肺炎克雷伯菌进行耐药机制研究和同源性分析。方法收集2012年3月上海新华医院临床微生物实验室从1例膀胱癌术后患者的血、尿、痰和盆腔引流液标本中分离得到耐碳青霉烯类抗生素肺炎克雷伯菌4株。分别采用:①改良Hodge试验筛选碳青霉烯酶;②PCR方法及基因测序检测耐药基因;③SDS-PAGE分析菌株外膜蛋白的改变等方法进行耐药机制研究。并采用ERIC-PCR方法进行DNA同源性分析。结果改良Hodge试验显示4株耐碳青霉烯类抗生素肺炎克雷伯菌均产碳青霉烯酶,通过PCR方法及基因测序确证皆产KPC-2酶。SDS-PAGE分析结果显示4株细菌的外膜蛋白表达不同于碳青霉烯类抗生素敏感型肺炎克雷伯菌。ERIC-PCR基因图谱显示此4株细菌的DNA指纹图谱完全一致。结论本研究中的4株耐碳青霉烯类抗生素肺炎克雷伯菌的主要耐药机制为产KPC-2酶以及外膜蛋白异常引起的外膜蛋白通透性改变,且这4株细菌为同一克隆株。
引用
收藏
页码:460 / 464
页数:5
相关论文
共 17 条
[1]  
Multicenter study using standardized protocols and reagents for evaluation of reproducibility of PCR-based fingerprinting of Acinetobacter spp. Crundmann HJ,Tower KJ,Kijkshoom L,et al. Journal of Clinical Microbiology . 1997
[2]  
Plasmid-mediated carbapenem-hydrolysingβ-lactamase,KPC-2,inKlebsiella pneumoniaeisolates. Smith Moland E,Hanson ND,Herrera VL,et al. J Anti microb Chemoth-er . 2003
[3]  
PCR detection of metallo-β-lactamase gene (blaIMP) in gram-negative rods resistant to broad-spectrum β-lactams. Kazuyoshi Senda,Yoshichika Arakawa,Satoshi Ichiyama,Kazumitsu Nakashima,Hideo Ito,Shinji Ohsuka,Kaoru Shimokata,Nobuo Kato,Michio Ohta. Journal of Clinical Microbiology . 1996
[4]  
Outbreak of infections caused by Pseudomonas aeruginosa producing VIM-carbapenemase in Greece. Tsakris A,Pournaras S,Woodford N,et al. Journal of Clinical Microbiology . 2000
[5]  
β-lactamase classification and amino acid sequences for TEM,SHV and OXA extended-spectrum and inhibitor resistant enzymes. Lahey Clinic. http://www.lahey.org/studies/webt.asp .
[6]  
Real-time PCR assayand a synthetic positive control for the rapid and sensitive de-tection of the emerging resistance gene New Delhi Metallo-β-lactamase-1(blaNDM-1). Krüttgen A,Razavi S,Imhl M,et al. Medical Microbiology and Immunology . 2011
[7]  
Emergence of oxacillinase-mediatedresistance to imipenem in Klebsiellapneumoniae. Poirel L,Héritier C,Tolün V,et al. Antimicrobial Agents and Chemotherapy . 2004
[8]  
In vivo transfer of plas-mid-encoded ACC-1AmpCfrom Klebsiella pneumoniae to Esche-richia coli in aninfant and selection of i mpermeability toi mipenem in K pneumoniae. Bidet P,,Burghoffer B,Gautier V,et al. Antimicrobial Agents and Chemotherapy . 2005
[9]  
Roles of β-lactamases and porins in activities of carbapenems and cephalosporins against Klebsiella pneumoniae. Martinez-Martinez L,Pascual A,Hernández-Allés S.et al. Antimicrobial Agents and Chemotherapy . 1999
[10]  
Ertapenem resistance among extended-spectrum-lactamase-producing Klebsiella pneumoniae isolates. Leavitt A,Chmelnitsky I,Colodner R,et al. Journal of Clinical Microbiology . 2009