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JAC Advance Access originally published online on April 7, 2009
Journal of Antimicrobial Chemotherapy 2009 63(6):1135-1141; doi:10.1093/jac/dkp122
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© The Author 2009. 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

Molecular epidemiology of Escherichia coli producing extended-spectrum β-lactamases in Lugo (Spain): dissemination of clone O25b:H4-ST131 producing CTX-M-15

Miguel Blanco1, Maria Pilar Alonso1,2, Marie-Hélène Nicolas-Chanoine3,4, Ghizlane Dahbi1, Azucena Mora1, Jesús E. Blanco1, Cecilia López1, Pilar Cortés5, Montserrat Llagostera5, Véronique Leflon-Guibout3, Beatriz Puentes1, Rosalía Mamani1, Alexandra Herrera1, María Amparo Coira2, Fernando García-Garrote2, Julia María Pita2 and Jorge Blanco1,*

1 Laboratorio de Referencia de E. coli, Departamento de Microbioloxía e Parasitolxía, Facultade de Veterinaria, Universidade de Santiago de Compostela, Lugo, Spain 2 Unidade de Microbioloxía Clínica, Complexo Hospitalario Xeral-Calde, Lugo, Spain 3 Service de Microbiologie, Hôpital AP-HP Beaujon, Clichy, France 4 Faculté de Médecine D. Diderot, Université Paris 7, Paris, France 5 Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Bellaterra (Barcelona), Spain

Received 30 December 2008; returned 13 February 2009; revised 27 February 2009; accepted 4 March 2009

* Corresponding author. Tel/Fax: +34-982-285936; E-mail: jorge.blanco{at}usc.es

Objectives: Having shown that the Xeral-Calde Hospital in Lugo (Spain) has been concerned by Escherichia coli clone O25:H4-ST131 producing CTX-M-15 (Nicolas-Chanoine et al. J Antimicrob Chemother 2008; 61: 273–81), the present study was carried out to evaluate the prevalence of this clone among the extended-spectrum β-lactamase (ESBL)-producing E. coli isolates and also to molecularly characterize the E. coli isolates producing ESBL other than CTX-M-15.

Methods: In the first part of this study, 105 ESBL-producing E. coli isolates (February 2006 to March 2007) were characterized with regard to ESBL enzymes, serotypes, virulence genes, phylogenetic groups, multilocus sequence typing (MLST) and PFGE. In the second part of this study, 249 ESBL-producing E. coli isolates (April 2007 to May 2008) were investigated only for the detection of clone O25b:H4-ST131 producing CTX-M-15 using a triplex PCR developed in this study and based on the detection of the new operon afa FM955459 [GenBank] and the targets rfbO25b and 3' end of the blaCTX-M-15 gene.

Results: Of the 105 ESBL-producing E. coli isolates, 60 (57.1%) were positive for CTX-M-14, 23 (21.9%) for CTX-M-15, 10 (9.5%) for SHV-12 and 7 (6.7%) for CTX-M-32. Serotypes, virulence genes, phylogenetic groups and molecular typing by PFGE demonstrated high homogeneity within those producing CTX-M-15 and high diversity within E. coli producing CTX-M-14 and other ESBLs. By PFGE, CTX-M-15-producing E. coli isolates O25b:H4 belonging to the phylogenetic group B2 and MLST profile ST131 were grouped in the same cluster. The epidemic strain of clone O25b:H4-ST131 represented 23.1%, 22.5% and 20.0% of all ESBL-producing E. coli isolated in 2006, 2007 and 2008, respectively.

Conclusions: CTX-M-type ESBLs, primarily CTX-M-14 and CTX-M-15, have emerged as the predominant types of ESBL produced by E. coli isolates in Lugo. In view of the reported findings, long-term care facilities for elderly people may represent a significant reservoir for E. coli clone O25b:H4-ST131 producing CTX-M-15. The triplex PCR developed in this work will be useful for rapid and simple detection of this clone.

Keywords: antibiotic resistance , CTX-M , E. coli , ESBLs


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