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Journal of Clinical Microbiology, June 2008, p. 2147-2148, Vol. 46, No. 6
0095-1137/08/$08.00+0 doi:10.1128/JCM.00427-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.
CTX-M-15-Producing Shigella sonnei Strain from a Czech Patient Who Traveled in Asia 

LETTER
International travel has been recognized as an important factor
in the diffusion of multiresistant bacteria, including organisms
that produce extended-spectrum β-lactamases (ESBLs) (
14,
15,
16). When followed by clonal spread and/or horizontal transfer
of ESBL-encoding plasmids in new areas, it may contribute to
the global "epidemiologic success" of particular ESBL types.
This is probably the case for some of the CTX-M enzymes (e.g.,
CTX-M-15), a family of derivatives of natural β-lactamases
of
Kluyvera spp. (
3,
12). Here we report the first identification
of ESBL-producing
Shigella sonnei in the Czech Republic, from
a 31-year-old female who traveled to China, Nepal, and India
in April and May 2006. Immediately after returning, she was
admitted with diarrhea and vomiting to a regional hospital.
S. sonnei was cultured from stool; the identification was performed
by ENTEROtest 24 (Pliva-Lachema Diagnostika, Brno, Czech Republic)
and
Shigella sonnei Polyvalent D (Denka Seiken, Tokyo, Japan).
The isolate was found to produce ESBL with the double-disk synergy
test (
10). MICs of various antimicrobials, determined by the
CLSI microdilution method (
5), showed a classical ESBL-mediated
pattern of β-lactam resistance, including cefotaxime and
ceftazidime (Table
1). The patient, not treated with antibiotics,
recovered in 21 days.
The isolate was subjected to mating with
Escherichia coli A15
Rif
r as previously described (
8); cefotaxime- and rifampin-resistant
transconjugants were obtained at a frequency of 10
–4 per
donor cell. Isoelectric focusing (
2) revealed that both the
S. sonnei isolate and its transconjugant expressed β-lactamases
with pIs of 8.9 and 5.4, of which the pI 8.9 enzyme hydrolyzed
cefotaxime and ceftazidime in the overlay assay (
2). PCR with
primers P1C and P2D, which are specific for
blaCTX-M-1-like
genes (
8), followed by DNA sequencing, identified the enzyme
as CTX-M-15, which is encoded by a gene identical to those originally
reported (
1,
11). Owing to the Glu240Gly mutation, CTX-M-15
belongs to the CTX-M enzymes with significant ceftazidime-hydrolyzing
activity (
7,
13). PCR and sequencing of the 5' flank of
blaCTX-M-15,
carried out as described previously (
1), showed an IS
Ecp1 element
placed 48 bp upstream from the gene. Such a genetic arrangement,
suggesting a single IS
Ecp1-mediated escape of
blaCTX-M-15 from
Kluyvera spp., has been found in enterobacteria from many countries
(
12), including India, where CTX-M-15 was identified originally
(
11). Plasmid replicon typing (
4), performed with the transconjugant's
DNA, revealed the presence of a sole FII replicon, characteristic
for plasmids that are associated with the worldwide spread of
blaCTX-M-15 genes (
6,
9). However, in contrast to many molecules
of that type (
6,
9), the plasmid did not transmit any other
resistance from the
S. sonnei isolate (Table
1) apart from that
to β-lactams (data not shown).
This report documents the repeated importation of a CTX-M-15 producer to Europe, where organisms with the same context of blaCTX-M-15, mostly E. coli, have already been disseminating (12), also in the Czech Republic (J.H., J.E., M.G., and P.U., unpublished data). It demonstrates further that the ongoing flow of ESBL-producing bacteria between different regions, including their constant supply from high-prevalence areas, makes the control of their wide spread enormously difficult.

ACKNOWLEDGMENTS
The study reported here was partially financed by research project
grant MSM 0021620819 from the Ministry of Education, Czech Republic.
J.E. and M.G. were partially supported by grant 6 PRCD LSHM-CT-2003-503-335
(COBRA) from the European Commission.

FOOTNOTES

Published ahead of print on 23 April 2008.


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Jaroslav Hrabák*
Department of Microbiology Faculty of Medicine in Plze Charles University in Prague Plze 305 99, Czech Republic
Joanna Empel
Marek Gniadkowski
Department of Molecular Microbiology National Medicines Institute Warsaw 00-725, Poland,1
Zbyn k Halbhuber
Karel Rébl
Department of Clinical Microbiology Institute of Health in Kolín Kladno 272 01, Czech Republic,2
Pavla Urbá ková
National Reference Laboratory for Antibiotics National Institute of Health Praha 100 42, Czech Republic,3
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* Phone: 420 377 40 29 32 Fax: 420 377 10 32 50 E-mail: Jaroslav.Hrabak{at}lfp.cuni.cz |
Journal of Clinical Microbiology, June 2008, p. 2147-2148, Vol. 46, No. 6
0095-1137/08/$08.00+0 doi:10.1128/JCM.00427-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.
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