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Antimicrobial Agents and Chemotherapy, August 2006, p. 2741-2750, Vol. 50, No. 8
0066-4804/06/$08.00+0     doi:10.1128/AAC.00274-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Dissemination and Persistence of blaCTX-M-9 Are Linked to Class 1 Integrons Containing CR1 Associated with Defective Transposon Derivatives from Tn402 Located in Early Antibiotic Resistance Plasmids of IncHI2, IncP1-{alpha}, and IncFI Groups

Ângela Novais,1 Rafael Cantón,1,2 Aránzazu Valverde,1 Elisabete Machado,1,3 Juan-Carlos Galán,1,2 Luísa Peixe,3 Alessandra Carattoli,4 Fernando Baquero,1,2 and Teresa M. Coque1,2*

Hospital Universitario Ramón y Cajal, IMSALUD, Madrid, Spain,1 Unidad de Resistencia a Antibióticos y Virulencia Bacteriana, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain,2 Faculdade de Farmácia, Universidade do Porto, Porto, Portugal;,3 Istituto Superiore di Sanità, Roma, Italy4

Received 3 March 2006/ Returned for modification 27 April 2006/ Accepted 6 June 2006

This study analyzes the diversity of In60, a class 1 integron bearing CR1 and containing blaCTX-M-9, and its association with Tn402, Tn21, and classical conjugative plasmids among 45 CTX-M-9-producing clinical strains (41 Escherichia coli strains, 2 Klebsiella pneumoniae strains, 1 Salmonella enterica strain, and 1 Enterobacter cloacae strain). Forty-five patients in a Spanish tertiary care hospital were studied (1996 to 2003). The diversity of In60 and association of In60 with Tn402 or mercury resistance transposons were investigated by overlapping PCR assays and/or hybridization. Plasmid characterization included comparison of restriction fragment length polymorphism patterns and determination of incompatibility group by PCR-based replicon typing, sequencing, and hybridization. CTX-M-9 plasmids belonged to IncHI2 (n = 26), IncP-1{alpha} (n = 10), IncFI (n = 4), and IncI (n = 1) groups. Genetic platforms containing blaCTX-M-9 were classified in six types in relation to the In60 backbone and in eight subtypes in relation to Tn402 derivatives. They were associated with Tn21 sequences when located in IncP-1{alpha} or IncHI2 plasmids. Our study identified blaCTX-M-9 in a high diversity of CR1-bearing class 1 integrons linked to different Tn402 derivatives, often to Tn21, highlighting the role of recombination events in the evolution of antibiotic resistance plasmids. The presence of blaCTX-M-9 on broad-host-range IncP-1{alpha} plasmids might contribute to its dissemination to hosts that were not members of the family Enterobacteriaceae.

* Corresponding author. Mailing address: Servicio de Microbiología, Hospital Universitario Ramón y Cajal, Carretera de Colmenar, km. 9.1, Madrid 28034, Spain. Phone: 34-91-336 83 30. Fax: 34-91-336 88 09. E-mail: mcoque.hrc{at}salud.madrid.org.

Antimicrobial Agents and Chemotherapy, August 2006, p. 2741-2750, Vol. 50, No. 8
0066-4804/06/$08.00+0     doi:10.1128/AAC.00274-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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