Nucleotide Sequence and Characterization of a Novel Cefotaxime-Hydrolyzing β-Lactamase (CTX-M-10) Isolated in Spain

Oliver, Antonio; Pérez-Díaz, J C; Coque, Teresa M.; Baquero, Fernando; Cantón, Rafael

doi:10.1128/aac.45.2.616-620.2001

Cited by 101 publications

(73 citation statements)

References 25 publications

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“…IEF and PCR results suggested that 10 of the 11 E. cloacae isolates harbored the same ESBL, which corresponds to the newly characterized CTX-M-10 enzyme in an E. coli isolate from our hospital (24). This ␤-lactamase has also been found in 40 and 10% of the ESBL-producing K. pneumoniae and E. coli clones recovered in different units from our hospital, respectively, suggesting that this ␤-lactamase is widespread in our institution (5).…”

Section: Discussionmentioning

confidence: 84%

Epidemiology of Extended-Spectrum β-Lactamase-Producing Enterobacter Isolates in a Spanish Hospital during a 12-Year Period

et al. 2002

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Fifteen Enterobacter clinical isolates (11 Enterobacter cloacae isolates, 3 Enterobacter aerogenes isolates, and 1 Enterobacter gergoviae isolate), representing 0.4% of all Enterobacter isolates recovered in our hospital from 1989 to 2000, were suspected of harboring an extended-spectrum ␤-lactamase (ESBL). These isolates were recovered from 14 different patients. ESBLs were transferred by conjugation into an Escherichia coli recipient strain. Pulsed-field gel electrophoresis (PFGE) revealed a single clone of E. aerogenes and six different clones of E. cloacae. Four of these E. cloacae clonal types were represented by only one isolate each, but the other two were represented by three and four isolates, respectively. Isoelectric focusing, susceptibility phenotyping, PCR analysis, and sequencing demonstrated the presence of three different ESBLs. The most frequent was the recently characterized CTX-M-10 ESBL, which was found in the E. gergoviae isolate and in all but one of the E. cloacae isolates. The remaining E. cloacae isolate harbored a TEM-27 ESBL, and the three E. aerogenes isolates harbored a TEM-24 ESBL. PFGE revealed that our E. aerogenes strain was indistinguishable from the French TEM-24-producing E. aerogenes endemic clone. Although a low prevalence of ESBL-producing Enterobacter isolates was found in our institution over a 12-year period, a diversity of nonepidemic E. cloacae clones was detected, as was the persistence of the CTX-M-10 ␤-lactamase. The presence of the TEM-24-producing E. aerogenes French clone in our institution also demonstrates the intercountry dissemination of ESBL-producing isolates.Plasmid-mediated extended-spectrum ␤-lactamases (ESBLs) were first reported in the mid-1980s (14, 15). Since then they have been responsible for several outbreaks caused by expanded-spectrum cephalosporin-resistant enterobacteria. Klebsiella pneumoniae and Escherichia coli are the most frequently involved organisms, and TEM-1-and SHV-1-derived ␤-lactamases are the most common ESBLs found in these species (25). These enzymes have also been reported at a much lower frequency among inducible AmpC ␤-lactamase-producing members of the family Enterobacteriaceae, such as Enterobacter spp., Citrobacter spp., Morganella morganii, and Serratia spp. (6,10,17,23,27,28,38). The most common ESBLs found in these species also belong to the TEM-and SHV-derived ␤-lactamases.Enterobacter species have been found to be one of the most important causes of nosocomial infections during the last few years (9, 30). These organisms have been associated with several outbreaks generally involving derepressed mutants overproducing their chromosomal ␤-lactamase or, more infrequently, expressing ESBL (7,10,23,30).The aims of this study were to establish the epidemiological relationship among all ESBL-producing Enterobacter isolates recovered in our institution over a decade, as well as to characterize the ESBLs expressed in these isolates and the plasmids harboring ESBL genes. In addition, we investigated if our ESBL-producing Entero...

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Section: Discussionmentioning

confidence: 84%

Epidemiology of Extended-Spectrum β-Lactamase-Producing Enterobacter Isolates in a Spanish Hospital during a 12-Year Period

et al. 2002

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“…The overall results showed a similarity between plasmids in strains 1, 4, 5, 7, 9, 10, 12, 13, and 14 (RFLP pattern I in Table 3) obtained from the 17 E. coli strains, thus suggesting identity in the plasmids harboring CTX-M-14 on these strains. However, strains 2, 3,6,8,11,15,16, and 17 did not show a similarity in their RFLP patterns, and their patterns were different from RFLP pattern I. Therefore, the CTX-M-14 gene was harbored in different plasmids on these strains, and so the possibility that a mobile genetic element might be involved in the dissemination of the CTX-M-14 gene in these plasmids cannot be ruled out.…”

Section: Selection Of Clinical Isolates and Patientsmentioning

confidence: 80%

Identification and Broad Dissemination of the CTX-M-14 β-Lactamase in Different Escherichia coli Strains in the Northwest Area of Spain

et al. 2002

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During the course of a molecular epidemiology study of mechanisms of antibiotic resistance in the area served by our hospital (516,000 inhabitants), we isolated the gene encoding CTX-M-14 ␤-lactamase. Thirty clinical strains (27 Escherichia coli and 3 Klebsiella pneumoniae isolates) with a phenotype of extended-spectrum ␤-lactamase were collected from January to October 2001 and studied for the presence of the CTX-M-14 ␤-lactamase gene. By isoelectric point determination, PCR, and nucleotide sequencing, we detected the presence of this gene in 17 E. coli strains belonging to 15 different genotypes (REP-PCR) causing infections in 17 different patients. Epidemiological studies based on medical records did not suggest any relationship between the patients infected with these E. coli strains and, interestingly, 7 of 30 patients harboring strains with extended-spectrum ␤-lactamases never had contact with the hospital environment before the clinical E. coli isolation. Conjugation experiments revealed that this gene was plasmid mediated in the 17 E. coli strains, and plasmid restriction fragment length polymorphisms showed 9 different patterns in the 17 E. coli strains. By PCR, the sequence of the tnpA transposase gene of the insert sequence ISEcp-1 was detected in all the plasmids harboring the CTX-M-14 gene. These results strongly suggest that plasmid dissemination between different E. coli strains in addition to a mobile element (transposon) around the ␤-lactamase gene may be involved in the spreading of the CTX-M-14 gene. This study reinforces the hypothesis that the epidemiology of the prevalence of the ␤-lactamase genes is changing and should alert the medical community to the increase in the emergence of the CTX-M ␤-lactamases worldwide.

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“…Because of their different enzymatic properties, ESBL producers vary in their capacities to hydrolyze specific ␤-lactam drugs. For example, organisms that produce TEM-10 and TEM-26, the most common ESBLs in the United States, have very high rates of resistance to ceftazidime and aztreonam, while the growing family of CA-sensitive CTX-M ␤-lactamase producers can hydrolyze cefotaxime at least 150 times more efficiently than ceftazidime (3,4,23). Confounding factors, such as the production of different ␤-lactamases by the same organism, can also lead to erroneous phenotypic conclusions.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Clinical Isolates of Enterobacteriaceae from Italy by the BD Phoenix Extended-Spectrum β-Lactamase Detection Method

et al. 2003

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Production of extended-spectrum ␤-lactamases (ESBLs) is an important mechanism of ␤-lactam resistance in Enterobacteriaceae. Identification of ESBLs based on phenotypic tests is the strategy most commonly used in clinical microbiology laboratories. The Phoenix ESBL test (BD Diagnostic Systems, Sparks, Md.) is a recently developed automated system for detection of ESBL-producing gram-negative bacteria. An algorithm based on phenotypic responses to a panel of cephalosporins (ceftazidime plus clavulanic acid, ceftazidime, cefotaxime plus clavulanic acid, cefpodoxime, and ceftriaxone plus clavulanic acid) was used to test 510 clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Providencia stuartii, Morganella morganii, Enterobacter aerogenes, Enterobacter cloacae, Serratia marcescens, Citrobacter freundii, and Citrobacter koseri. Of these isolates, 319 were identified as ESBL producers, and the remaining 191 were identified as non-ESBL producers based on the results of current phenotypic tests. Combined use of isoelectric focusing, PCR, and/or DNA sequencing demonstrated that 288 isolates possessed bla TEM-1 -and/or bla SHV-1 -derived genes, and 28 had a bla CTX-M gene. Among the 191 non-ESBL-producing isolates, 77 isolates produced an AmpC-type enzyme, 110 isolates possessed TEM-1, TEM-2, or SHV-1 ␤-lactamases, and the remaining four isolates (all K. oxytoca strains) hyperproduced K1 chromosomal ␤-lactamase. The Phoenix ESBL test system gave positive results for all the 319 ESBL-producing isolates and also for two of the four K1-hyperproducing isolates of K. oxytoca. Compared with the phenotypic tests and molecular analyses, the Phoenix system displayed 100% sensitivity and 98.9% specificity. These findings suggest that the Phoenix ESBL test can be a rapid and reliable method for laboratory detection of ESBL resistance in gram-negative bacteria.

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Nucleotide Sequence and Characterization of a Novel Cefotaxime-Hydrolyzing β-Lactamase (CTX-M-10) Isolated in Spain

Cited by 101 publications

References 25 publications

Epidemiology of Extended-Spectrum β-Lactamase-Producing Enterobacter Isolates in a Spanish Hospital during a 12-Year Period

Epidemiology of Extended-Spectrum β-Lactamase-Producing Enterobacter Isolates in a Spanish Hospital during a 12-Year Period

Identification and Broad Dissemination of the CTX-M-14 β-Lactamase in Different Escherichia coli Strains in the Northwest Area of Spain

Characterization of Clinical Isolates of Enterobacteriaceae from Italy by the BD Phoenix Extended-Spectrum β-Lactamase Detection Method

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