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Journal of Clinical Microbiology, December 2006, p. 4309-4315, Vol. 44, No. 12
0095-1137/06/$08.00+0     doi:10.1128/JCM.00817-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Establishing Clonal Relationships between VIM-1-Like Metallo-ß-Lactamase-Producing Pseudomonas aeruginosa Strains from Four European Countries by Multilocus Sequence Typing{triangledown}

Christian G. Giske,1,{dagger}* Balázs Libisch,2,{dagger}* Céline Colinon,3,{ddagger} Effie Scoulica,4 Laura Pagani,5 Miklós Füzi,2 Göran Kronvall,1 and Gian Maria Rossolini3

Department of Clinical Microbiology L2:02, Karolinska Institutet-MTC, Karolinska University Hospital Solna, SE-17176 Stockholm, Sweden,1 Department of Bacteriology, National Center for Epidemiology, Gyáli út 2-6, 1097 Budapest, Hungary,2 Department of Molecular Biology, Section of Microbiology, University of Siena, I-53100 Siena, Italy,3 Department of Clinical Bacteriology, Parasitology, Zoonoses and Geographical Medicine, Faculty of Medicine, University of Crete, 71409 Heraklion, Greece,4 Department of Morphology, Imaging and Clinical Sciences, Section of Microbiology, University of Pavia, I-27100 Pavia, Italy5

Received 17 April 2006/ Returned for modification 12 July 2006/ Accepted 21 September 2006

Ten multidrug-resistant Pseudomonas aeruginosa strains producing VIM-1-like acquired metallo-ß-lactamases (MBLs), isolated from four European countries (Greece, Hungary, Italy, and Sweden), were analyzed for genetic relatedness by several methodologies, including fliC sequence analysis, macrorestriction profiling of genomic DNA by pulsed-field gel electrophoresis (PFGE), random amplification of polymorphic DNA (RAPD), and multilocus sequence typing (MLST). The four approaches yielded consistent results overall but showed different resolution powers in establishing relatedness between isolates (PFGE > RAPD > MLST > fliC typing) and could usefully complement each other to address issues in the molecular epidemiology of P. aeruginosa strains producing acquired MBLs. In particular, the recently developed MLST approach was useful in revealing clonal relatedness between isolates when this was not readily apparent using RAPD and PFGE, and it suggested a common ancestry for some of the VIM-1-like MBL-positive P. aeruginosa strains currently spreading in Europe. The MBL producers belonged in three clonal complexes/burst groups (BGs). Of these, one corresponded to the previously described BG4 and included serotype O12 strains from Hungary and Sweden, while the other two were novel and included serotype O11 or nonserotypable strains from Greece, Sweden, and/or Italy. Comparison of the integrons carrying blaVIM-1-like cassettes of various isolates revealed a remarkable structural heterogeneity, suggesting the possibility that multiple independent events of acquisition of different blaVIM-containing integrons had occurred in members of the same clonal lineage, although a contribution of integrase-mediated cassette shuffling or other recombination mechanisms during the evolution of similar strains could also have played a role in determining this variability.

* Corresponding author. Mailing address for Christian G. Giske: Department of Clinical Microbiology L2:02, Karolinska Institutet-MTC, Karolinska University Hospital Solna, SE-17176 Stockholm, Sweden. Phone: 46 8 517 73574. Fax: 46 8 30 80 99. E-mail: christian.giske{at}karolinska.se. Mailing address for Balázs Libisch: Department of Bacteriology, National Center for Epidemiology, Gyáli út 2-6, 1097 Budapest, Hungary. Phone: 36 1 476 1118. Fax: 36 1 476 1234. E-mail: libischb{at}oek.antsz.hu.

{triangledown} Published ahead of print on 4 October 2006.

{dagger} C.G.G. and B.L. contributed equally to this paper.

{ddagger} Present address: Laboratory of Radioecology and Ecotoxicology, Institute for Radioprotection and Nuclear Safety, Cadarache, 13115 Saint Paul Lez Durance, France.

Journal of Clinical Microbiology, December 2006, p. 4309-4315, Vol. 44, No. 12
0095-1137/06/$08.00+0     doi:10.1128/JCM.00817-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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