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JAC Advance Access originally published online on November 6, 2008
Journal of Antimicrobial Chemotherapy 2009 63(1):60-66; doi:10.1093/jac/dkn453
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© The Author 2008. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Original research

Characterization of plasmids encoding blaESBL and surrounding genes in Spanish clinical isolates of Escherichia coli and Klebsiella pneumoniae

Karol Diestra1,2, Carlos Juan3, Tânia Curiao4,5, Bartolomé Moyá3, Elisenda Miró1, Jesús Oteo6, Teresa M. Coque4,5,7, María Pérez-Vázquez6, José Campos6, Rafael Cantón4,5,7, Antonio Oliver3, Ferran Navarro1,2,* on behalf of Red Española de Investigación en Patología Infecciosa (REIPI), Spain

1 Servicio de Microbiología, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain 2 Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, Cerdanyola del Vallès, Spain 3 Servicio de Microbiología, Hospital Son Dureta, Palma de Mallorca, Spain 4 Servicio de Microbiología, Hospital Universitario Ramón y Cajal, Madrid, Spain 5 CIBER en Epidemiología y Salud Pública (CIBERESP), Spain 6 Laboratorio de Antibióticos, Servicio de Bacteriología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain 7 Unidad de Resistencia a Antibióticos y Virulencia Bacteriana Asociada al Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain

Received 22 August 2008; returned 15 September 2008; revised 3 October 2008; accepted 7 October 2008

* Correspondence address. Servicio de Microbiología, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain. Tel: +34-932919071; Fax: +34-932919070; E-mail: fnavarror{at}santpau.cat

Objectives: The aim of the study was to characterize plasmids that harbour blaESBL genes and their genetic environment in Escherichia coli and Klebsiella pneumoniae clones circulating in Spain.

Methods: The incompatibility group of plasmids within 58 strains harbouring blaCTX-M (n = 45) and blaSHV (n = 15) genes was determined by rep-typing-PCR and hybridization. The blaESBL genetic environment was determined by PCR and sequencing.

Results: The blaCTX-M-9 genes (n = 14) were linked to In60 located in IncI1 (50%) or IncHI2 plasmids (28%). All blaCTX-M-14 genes (n = 13) were flanked by ISEcp1 and IS903 and 12 were associated with IncK plasmids. One of two blaCTX-M-10 genes was present in an IncK plasmid, but both genes were linked to a phage-related element. Five of seven blaCTX-M-1 (71%), all three blaCTX-M-32 and one of two blaCTX-M-3 genes were linked to IncN plasmids. The other blaCTX-M-3 gene was linked to IncA/C and the remaining two blaCTX-M-1 genes to IncFII plasmids. Three blaCTX-M-15 genes were associated with IncF (repFIA) and one with IncFII plasmids. All these genes from blaCTX-M group-1 showed the ISEcp1 upstream truncated by different insertion sequences. Forty-three percent of blaSHV-12 genes (n = 14) were located in IncI1 plasmids, all flanked by the IS26 and DEOR region. The only detected blaSHV-5 gene was located in an IncFII plasmid and flanked by recF and DEOR regions.

Conclusions: A diversity of the plasmid incompatibility groups that harbour blaESBL genes was observed, except for the blaCTX-M-14 gene. Moreover, a high variability was confirmed in the genetic environment of these genes as a result of insertion and deletion events.

Keywords: ESBL-producing , incompatibility group , antimicrobial resistance surveillance , mechanisms of resistance


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