Characterization of Resistance Genes and Polymerase Chain Reaction-Based Replicon Typing in Carbapenem-Resistant Klebsiella pneumoniae

Microb Drug Resist. 2019 May;25(4):551-557. doi: 10.1089/mdr.2018.0231. Epub 2018 Nov 20.

Abstract

Background: Fifty isolates of Klebsiella pneumoniae isolated from clinical samples between 2012 and 2016 that were found to be resistant to carbapenems were included in this study. Materials and Methods: Resistance genes were investigated by performing PCR. Plasmid typing was performed using PCR-based replicon typing. The clonal relationships between the strains were investigated using pulsed-field gel electrophoresis (PFGE). Results: OXA-48-type carbapenemase genes were detected in 86% (n = 43/50) of K. pneumoniae isolates, whereas NDM-type carbapenemase genes were detected in 14% (n = 7/50) of the isolates. blaTEM was detected 60% (n = 30) of the strains, blaSHV in 78% (n = 39), blaCTX-M-1 in 48% (n = 24), and blaCTX-M-2-type β-lactamase in 10% (n = 5). blaCTX-M-1 and blaSHV were concomitantly distributed in 40% (n = 20) of the strains, blaTEM and blaSHV in 54% (n = 27), blaTEM, blaSHV, and blaCTX-M-1 in 32% (n = 16) and blaCTX-M-1 and blaCTX-M-2 in 10% (n = 5). Strain numbers 66, 69, 76, 77, and 78 coproduced carbapenemases, blaCTX-M-1 and blaCTX-M-2 in addition to blaOXA-48 or blaNDM-1 that were described as hybrid strains. IncR-type replicon was found in 50% (n = 25) of 50 isolates with plasmid typing, whereas IncA/C-type replicon was detected in 40% (n = 20) and IncFIIK-type replicon in 18% (n = 9) of the isolates. Outcomes of the transformation experiments showed that the OXA-48 gene was carried to the receiver cell on FII plasmids. No dominant epidemic clone was detected through PFGE. Conclusion: OXA-48 carbapenemase was found to be the most prevalent type of enzyme in our hospital, and the presence of NDM-1-type carbapenemase-carrying strain and an increase in their rate were detected.

Keywords: Enterobacteriaceae (CRE); NDM-1; OXA-48; carbapenemase.

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Bacterial Proteins / genetics
  • Carbapenems / pharmacology*
  • Drug Resistance, Bacterial / genetics*
  • Humans
  • Klebsiella Infections / drug therapy
  • Klebsiella Infections / microbiology
  • Klebsiella pneumoniae / genetics*
  • Microbial Sensitivity Tests / methods
  • Multilocus Sequence Typing / methods
  • Plasmids / genetics
  • Polymerase Chain Reaction / methods
  • Replicon / genetics*
  • beta-Lactamases / genetics

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Carbapenems
  • beta-lactamase CTX-2
  • beta-lactamase TEM-3
  • beta-Lactamases
  • carbapenemase