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JAC Advance Access published online on July 30, 2008

Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkn287
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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

Expansion and countrywide dissemination of ST11, ST15 and ST147 ciprofloxacin-resistant CTX-M-15-type β-lactamase-producing Klebsiella pneumoniae epidemic clones in Hungary in 2005—the new ‘MRSAs’?

Ivelina Damjanova1,*, Ákos Tóth2, Judit Pászti1, Gabriella Hajbel-Vékony1, Melinda Jakab1, Judit Berta2, Hedda Milch1 and Miklós Füzi2

1 Department of Phage Typing and Molecular Epidemiology, National Center for Epidemiology, Gyáli út 2-6, 1097 Budapest, Hungary 2 Department of Bacteriology, National Center for Epidemiology, Gyáli út 2-6, 1097 Budapest, Hungary

Received 7 February 2008; returned 31 March 2008; revised 13 June 2008; accepted 20 June 2008


* Corresponding author. Tel: +36-1-4761265; Fax: +36-1-4761234; E-mail: damjanova.ivelina{at}oek.antsz.hu

Objectives: To investigate the molecular epidemiology of ciprofloxacin-resistant CTX-M-15-producing Klebsiella pneumoniae epidemic clones (ECs) isolated from six nosocomial outbreaks and sporadic cases during 2005 in Hungary.

Methods: Two hundred and eighty-one extended-spectrum β-lactamase (ESBL)-producing K. pneumoniae clinical isolates collected from 41 centres were submitted to the National ESBL Reference Laboratory for further investigations. Of the 281 strains, 75 isolates proved to be SHV producers, whereas 6 isolates were ciprofloxacin-susceptible CTX-M-type ESBL producers. One hundred and ninety-six ciprofloxacin-resistant CTX-M-type β-lactamase-producing isolates collected from 35 centres were subjected to macrorestriction profile analysis. Furthermore, molecular typing was performed by PCR and sequencing of several antibiotic resistance genes, plasmid profile analysis, transfer of resistance determinants and multilocus sequence typing (MLST).

Results: PFGE revealed the existence of three genetic clusters defined as ECs, where 129 isolates belonged to the previously described Hungarian EC (HEC), 46 isolates to epidemic clone II (EC II) and 21 isolates to epidemic clone III (EC III), respectively. All isolates harboured plasmids ranging from 2.0 to 230 kb. PstI digestion of plasmid DNA from transconjugants/transformants revealed diverse restriction patterns from distinct ECs. Sequence analysis of β-lactamase genes from 19 selected isolates detected blaCTX-M-15 and blaOXA-1 in strains from all three ECs and blaTEM-1 in EC III isolates located on large plasmids. ISEcpI associated with CTX-M-15 was detected only on a 50 kb non-conjugative plasmid from EC III. MLST identified three allelic profiles: ST 15 (HEC), ST 11 (EC III) and the novel ST 147 (EC II), which correspond to the PFGE clusters, respectively.

Conclusions: In 2005, 97% of all CTX-M-producing K. pneumoniae isolates detected across Hungary were highly ciprofloxacin-resistant CTX-M-15 producers and represented just three stable genetic clones.

Key Words: aac(6')-Ib-cr , ISEcp1 , MLST


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