Molecular characterization of carbapenem-resistant Klebsiella pneumoniae isolates from a university hospital in Brazil

  • Ana Carolina Polano Vivan University of the Sacred Heart, Bauru, Brazil
  • Juliana Ferraz Rosa São Paulo University, São Paulo, Brazil
  • Camila Fonseca Rizek São Paulo University, São Paulo, Brazil
  • Marsileni Pelisson Londrina State University, Londrina, Brazil
  • Silvia Figueiredo Costa São Paulo University, São Paulo, Brazil
  • Mariangela Hungria Embrapa Soja, Londrina, PR, Brazil
  • Renata Kobayashi Londrina State University, Londrina, Brazil
  • Eliana Carolina Vespero Londrina State University, Londrina, Brazil
Keywords: Klebsiella pneumoniae, carbapenemase, KPC, carbapenem resistance

Abstract

Introduction: The emergence of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae (KPC-Kpn) isolates is attracting significant attention in nosocomial infection settings. K. pneumoniae is the main pathogen that harbours blaKPC genes.

Methodology: This study evaluated 54 K. pneumoniae carbapenem-resistant isolates from patients hospitalized at the University Hospital of Londrina, between July 2009 and July 2010. The isolates were phenotypically screened for carbapenemase production and submitted for genotypic confirmation by polymerase chain reaction (PCR) for KPC, metallo-β-lactamases, OXA-48, and extended-spectrum beta-lactamase genes. The absence of outer membrane proteins (OMP) was investigated by SDS-PAGE. The susceptibility profile was determined by broth microdilution, according to Clinical and Laboratory Standards Institute protocol.

Results: All isolates were phenotypically positive for class A carbapenemase production, but negative for metallo-β-lactamase activity. PCR analysis demonstrated that all isolates carried blaKPC genes and sequencing showed that all strains belonged to KPC-2 subtype. Four strains did not show porin expression, and all isolates were resistant to ertapenem, meropenem, and imipenem. Susceptibility rates reached 35.2% for gentamicin, 85.2% for polymixyn B, 87% for colistin, and 98.1% for both tigecycline and fosfomycin. Pulsed-field gel electrophoresis showed six clones, and three of them predominated among the isolates.

Conclusions: KPC-2-producing K. pneumoniae is becoming predominant among carbapenem-resistant K. pneumoniae isolates at the hospital. The association of the enzyme KPC with other resistance determinants, such as loss of porins, may increase the severity of the situation of nosocomial infections. There is an urgent need to develop strategies for infection control and prevention.

Author Biographies

Ana Carolina Polano Vivan, University of the Sacred Heart, Bauru, Brazil

Department of Health Sciences

Juliana Ferraz Rosa, São Paulo University, São Paulo, Brazil

Laboratory of Bacteriology (LIM-54), Medical School of the University of São Paulo

Camila Fonseca Rizek, São Paulo University, São Paulo, Brazil

Laboratory of Bacteriology (LIM-54), Medical School of the University of São Paulo

Marsileni Pelisson, Londrina State University, Londrina, Brazil

Department of Pathology, Clinical Analysis and Toxicological Center - Health Sciences - 

Silvia Figueiredo Costa, São Paulo University, São Paulo, Brazil

Laboratory of Bacteriology (LIM-54), Hospital das Clínicas, Medical School of the University of São Paulo, São Paulo, SP, Brazil

Mariangela Hungria, Embrapa Soja, Londrina, PR, Brazil

Embrapa Soja, Londrina, PR, Brazil

Renata Kobayashi, Londrina State University, Londrina, Brazil

Department of Microbiology, Center of Biological Sciences, State University of Londrina Londrina, Paraná, Brazil

Eliana Carolina Vespero, Londrina State University, Londrina, Brazil

Department of Pathology, Clinical Analysis and Toxicological Center - Health Sciences - State University of Londrina, Londrina, PR, Brazil

Published
2017-06-01
How to Cite
1.
Vivan ACP, Rosa JF, Rizek CF, Pelisson M, Costa SF, Hungria M, Kobayashi R, Vespero EC (2017) Molecular characterization of carbapenem-resistant Klebsiella pneumoniae isolates from a university hospital in Brazil. J Infect Dev Ctries 11:379-386. doi: 10.3855/jidc.8614
Section
Original Articles