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Transconjugation and genotyping of the plasmid-mediated AmpC β-lactamase and extended-spectrum β-lactamase genes in Klebsiella pneumoniae

SHI Wei-feng ZHOU Jun QIN Jian-ping

中华医学杂志(英文版)2009,Vol.122Issue(9):1092-1096,5.
中华医学杂志(英文版)2009,Vol.122Issue(9):1092-1096,5.DOI:10.3760/cma.j.issn.0366-6999.2009.09.015

Transconjugation and genotyping of the plasmid-mediated AmpC β-lactamase and extended-spectrum β-lactamase genes in Klebsiella pneumoniae

Transconjugation and genotyping of the plasmid-mediated AmpC β-lactamase and extended-spectrum β-lactamase genes in Klebsiella pneumoniae

SHI Wei-feng 1ZHOU Jun 2QIN Jian-ping3

作者信息

  • 1. Department of Clinical Laboratory, Third Affiliated Hospital of Suzhou University, Changzhou, Jiangsu 213003, China
  • 2. Department of Respiratory Medicine, Third Affiliated Hospital of Suzhou University, Changzhou, Jiangsu 213003, China
  • 3. Department of Clinical Laboratory, Changzhou Traditional Chinese Medicine Hospital, Changzhou, Jiangsu 213003, China
  • 折叠

摘要

Abstract

Backgroud AmpC β-lactamases and extended-spectrum β-lactamases (ESBLs) are becoming predominant causes of resistance to third and forth-generation cephalosporins in Klebsiella pneumoniae (K. Pneumoniae). It is very difficult to treat infectious diseases caused by multidrug-resistant K. Pneumoniae. The purpose of the present study was to investigate transconjugation and characteristics of β-lactamase genes in K. Pneumoniae producing AmpC β-lactamases and ESBLs.Methods AmpC β-lactamases were detected by three-dimension test and ESBLs by disc confirmatory test. Minimum inhibitory concentrations (MICs) were determined by agar dilution. Transfer of resistance to EC600 (Rif') was attempted by conjugation in broth and screened on agar containing cefotaxime (2 μg/ml) plus rifampin (1024 μg/ml). The genes encoding AmpC or ESBLs and their transconjugants were detected by PCR and verified by DNA sequencing. Results The resistant rates to ampicillin and piperacillin were 100% in 18 isolates of K. Pneumoniae. However, imipenem was still of great bactericidal activity on K. Pneumoniae, and its MIC50 was 0.5 μg/mL. Eleven β-lactamase genes, including TEM-1, TEM-11, SHV-13, SHV-28, CTX-M-9, CTX-M-22, CTX-M-55, OXA-1, LEN, OKP-6 and DHA-1, were found from 18 isolates. And at least one β-lactamase gene occurred in each isolate. To our surprise, there were six β-lactamase genes in the CZ04 strain. Among 18 isolates of K. Pneumoniae, the partial resistant genes in 8 isolates were conjugated successfully, which had 100% homological sequence with donors by sequence analysis. Compared with donors, 8 transconjugants had attained resistance to most β-lactams, including ampicillin, piperacillin, cefoxitin, cefotaxime and aztreonam, or even amikacin and gentamicin.Conclusions R plasmids can be easily transferred between the resistant and sensitive negative bacilli. It is very difficult to block and prevent the spread of antimicrobial resistance. So more attention should be paid to reducing the frequency, times and dosage of antimicrobials, especially third or fourth cephalosporins.

关键词

Klebsiella pneumoniae/plasmid/transconjugation/β-lactamase

Key words

Klebsiella pneumoniae/plasmid/transconjugation/β-lactamase

分类

医药卫生

引用本文复制引用

SHI Wei-feng,ZHOU Jun,QIN Jian-ping..Transconjugation and genotyping of the plasmid-mediated AmpC β-lactamase and extended-spectrum β-lactamase genes in Klebsiella pneumoniae[J].中华医学杂志(英文版),2009,122(9):1092-1096,5.

中华医学杂志(英文版)

OACSTPCDMEDLINESCI

0366-6999

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