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Antimicrobial Agents and Chemotherapy, November 2000, p. 3239-3240, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
LETTERS TO THE EDITOR
First Isolation of a CTX-M-3-Producing Enterobacter
cloacae in France
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LETTER |
At the beginning of the 1990s, a new class A extended-spectrum
-lactamase (ESBL), MEN-1 (CTX-M-1), was characterized in
Escherichia coli strains isolated from Italian and
German patients (1, 2). CTX-M-1 was the first member
of the CTX-M -lactamase family, which now comprises nine members:
CTX-M-1 (MEN-1) (1, 2), CTX-M-2 (2), Toho-1
(7), CTX-M-3 (6), CTX-M-4 (6), CTX-M-5
(4), Toho-2 (8), CTX-M-6 (5), CTX-M-7
(5), and CTX-M-8 (3). These ESBLs conferred
higher cefotaxime MICs than those of ceftazidime.
E. coli (1, 7-10) and Salmonella
typhimurium (2, 5, 6, 11) strains are the species most
frequently reported to produce CTX-M enzymes. These enzymes are
reported mainly in three geographic areas: South America (2,
3), East Europe (4-7, 10, 11), and Japan (8,
9).
During a multicenter survey of ESBLs in France in 1998 (C. De
Champs, D. Sirot, C. Chanal, J. Sirot, and the French Study Group,
Abstr. 39th Intersci. Conf. Antimicrob. Agents Chemother., abstr.
149.C2-1485, p. 169, 1999), an Enterobacter cloacae strain (Ver-1) was selected for its resistance to broad-spectrum
cephalosporins and a positive double-disk synergy test. This strain was
isolated from a 56-year-old man admitted to the Versailles Hospital (Le Chesnay, France) with unbalanced non-insulin-dependent diabetes and
severe arteriopathy. His last hospitalization was in 1996 for
noninfected urinary tract retention. He had osteitis and a foot
ulcer infected by a Staphylococcus aureus strain. After 2 days of treatment with oxacillin, the E. cloacae strain,
Ver-1, was isolated from a urine sample collected for dysuria. This
colonization was not treated.
Ver-1 was also resistant to tetracycline, co-trimoxazole, gentamicin,
and tobramycin. The ESBL phenotype was transferred to E. coli HB101, resistant to rifampin, at 37°C during an overnight mating assay on solid Mueller-Hinton medium containing rifampin (300 µg/ml). The E. coli transconjugant, designated TrVer-1,
did not exhibit cotransferred resistance markers.
Table 1 shows the MICs of -lactams,
determined by the agar dilution method, for the strain E. cloacae Ver-1 and its E. coli transconjugant
TrVer-1. These two strains were resistant to penicillins. MICs of
cefotaxime (128 to 32 µg/ml) were 16- to 64-fold higher than those of
ceftazidime (8 to 0.5 µg/ml). The -lactam inhibitors clavulanate
and tazobactam restored partially or totally the susceptibilities to
piperacillin and cephalosporins.
View this table:
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TABLE 1.
MICs of -lactams for E. cloacae isolate
Ver-1 and its E. coli transconjugant TrVer-1 in comparison
with wild-type E. cloacae and TEM-1-producing E.
colia
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|
Sonicates of the clinical strain and its E. coli
transconjugant were subjected to analytical isoelectric focusing over
the pH range of 3 to 10. Both E. cloacae and its E. coli transconjugant produced a -lactamase of isoelectric point
8.4, associated with a -lactamase of pI 5.4.
PCR and direct DNA sequencing identified the -lactamase of pI 5.4 as
TEM-1 penicillinase. No PCR products were obtained with primers
specific for blaSHV. In contrast, positive
amplification was obtained with primers CTX-M-3A
(5'-GGTTAAAAAATCGCG-3') and CTX-M-3B
(5'-TTACAAACCGTCGGTGA-3'), which amplified the complete sequence of open reading frame blaCTX-M-3. The
obtained DNA sequence of the PCR products exhibited 100% identity to
the sequence blaCTX-M-3 (8).
While only CTX-M-1 and -2 were characterized in 1990 and 1992 (1,
2), seven new CTX-M enzymes were described in 1998 and 1999, showing that the CTX-M family of ESBLs was small but rapidly growing.
CTX-M-3 was first characterized in 1998 for Citrobacter freundii and E. coli strains at Praski Hospital in
Poland (8). This enzyme has spread in other species of the
Enterobacteriaceae (Klebsiella pneumoniae,
Klebsiella oxytoca, E. cloacae, and
Morganella morganii) and was the more frequently observed
ESBL (11). Here, we report the first characterization of a
CTX-M-3-producing strain isolated from a French patient with no history
of travel the year before. The spread of the CTX-M-3 enzyme from East
European countries cannot be excluded. Tassios et al. (11)
have shown the probable spread of a CTX-M-4-producing S. typhimurium clone in Russia, Greece, and Hungary. However, the
emergence of CTX-M enzymes from widespread environmental bacteria
could also explain their spread.
During the study of ESBLs (De Champs et al., 39th ICAAC) which led to
characterization of these CTX-M-3-producing E. cloacae strains, 79 ESBL-producing strains were isolated. Only one
CTX-M-producing strain was observed. Thus, this
CTX-M-3-producing strain seems to be a sporadic
isolate. However, in view of the spread of CTX-M-producing strains in East European countries, emergence of CTX-M-producing strains could be observed in France. The characterization of this CTX-M-3-producing strain highlights the feasibility of this process and
constitutes a forewarning of the probable existence of CTX-M-producing strains in France.
| |
FOOTNOTES |
*
Phone: 33 1 39 63 93 75
Fax: 33 1 39 63 93 12
E-mail: fdp{at}fc.horus-medical.fr
| |
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| | | | |
Florence Doucet-Populaire*
J. C. Ghnassia
Service de Microbiologie
Centre Hospitalier de Versailles
177 rue de Versailles
78157 Le Chesnay
France
|
| | | | |
R. Bonnet
J. Sirot
Laboratoire de Bacteriologie
Faculté de Médecine
63001 Clermont-Ferrand
France
|
Antimicrobial Agents and Chemotherapy, November 2000, p. 3239-3240, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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