AAC
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Conceição, T.
Right arrow Articles by Barros, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Conceição, T.
Right arrow Articles by Barros, R.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, January 2005, p. 476, Vol. 49, No. 1
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.1.476.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

LETTER TO THE EDITOR

First Isolation of blaVIM-2 in Klebsiella oxytoca Clinical Isolates from Portugal


   LETTER
Top
 Letter
References
 
VIM-type carbapenemases were originally detected in Europe (2, 4, 7) and have been essentially found in Pseudomonas aeruginosa, as well as in Enterobacteriaceae (2, 6, 8). The blaVIM genes are often carried by mobile gene cassettes inserted into class 1 integrons (7, 11). The Klebsiella genus is responsible for the most frequent human nosocomial infections of the respiratory and urinary tracts. Four Klebsiella oxytoca clinical isolates were recovered by blood culture from neonatal patients at a pediatric hospital. The clinical isolates showed resistance to amoxicillin and ticarcillin, which was restored by clavulanate, aminoglycosides, and fluoroquinolones, and intermediate susceptibility to imipenem (MIC, 4 µg/ml) and broad-spectrum cephalosporins and aztreonam (MICs, 8 and 16 µg/ml) and were susceptible to meropenem (MIC, 0.075 µg/ml). A deformation of ellipses between the two gradient sections with E-test MBL (metallo-ß-lactamase) strips was indicative of MBL production (10).

Plasmid DNA was prepared by the alkaline lysis method (9), and the same 50-kb plasmid was detected in all of the isolates. Conjugation experiments were attempted with rifampin-resistant strain Escherichia coli C600 in liquid and solid media. E. coli DH10B was used as the bacterial host in electroporation experiments. No resistance phenotype transfer was observed in conjugation or transformation experiments.

M13 PCR fingerprinting, used as a typing method, was performed by using the core region of bacteriophage M13 as the primer (3). The four isolates have the same M13 profile.

Since several ß-lactamase genes are part of gene cassettes included in class 1 integrons, PCR was performed for class 1 integrons with 5'-CS and 3'-CS primers (5) on plasmid DNA from K. oxytoca 17FFUL purified by a QIAGEN Plasmid Midi Kit, yielding an amplicon with a size of 3,500 bp. Consecutive PCR amplifications with specific primers for MBL and aminoglycoside acetyltransferase genes were performed. Analysis of sequence data revealed the presence of four gene cassettes in the class 1 integron. The first cassette contained the blaVIM-2 gene, coding for an MBL. The second and the third cassettes contained genes coding for 6'-N-aminoglycoside acetyltransferase (aacA4) and an aminoglycoside adenylyltransferase (aadA1), respectively. The last gene cassette contains a blaOXA-2 gene coding for an oxacillinase.

As in P. aeruginosa COL-1 (7), the blaVIM-2 gene was found to be carried on a plasmid-borne integron. However, the VIM-2-encoding plasmids and the structure of the blaVIM-2 gene cassette were different in the two bacteria. Key signatures of class 1 integrons were identified, such as a 5'-CS attI1 recombination site, a core site (GTTATGC), and an inverse core site (GCATAAC). The 59-base element was 72 bp long as the 59-base element of the blaVIM-2 gene cassette in COL-1 but differed by three nucleotides, showing 95.8% homology.

A chromosomal blaVIM-2 gene in a P. aeruginosa clinical isolate was already detected in Portugal (1), and the detection of a plasmid-borne blaVIM-2 gene in K. oxytoca demonstrates that the problem of MBL-producing pathogens no longer involves gram-negative nonfermenters alone but also involves enterobacteria.

This study found that a clonal VIM-2 spread occurred in the pediatric ward during the study.


   REFERENCES
Top
 Letter
 References
 

  1. Cardoso, O., R. Leitão, A. Figueiredo, J. Sousa, A. Duarte, and L. Peixe. 2002. Metallo-ß-lactamase VIM-2 in clinical isolates of Pseudomonas aeruginosa from Portugal. Microb. Drug Resist. 8(2):93-97.[CrossRef][Medline]
  2. Giakkoupi, P., A. Xanthaki, M. Kanelopoulou, A. Vlahaki, V. Miriagou, S. Kontou, E. Papafraggas, H. Malamou-Lada, L. S. Tzouvelekis, N. J. Legakis, and A. C. Vatopoulos. 2003. VIM-1 metallo-ß-lactamase-producing Klebsiella pneumoniae strains in Greek hospitals. J. Clin. Microbiol. 41:3893-3896.[Abstract/Free Full Text]
  3. Grundmann, H. J., K. J. Towner, L. Dijkshoorn, P. Gerner-Smidt, M. Maher, H. Seifert, and M. Vaneechoutte. 1997. Multicenter study using standardized protocols and reagents for evaluation of reproducibility of PCR-based fingerprinting of Acinetobacter spp. J. Clin. Microbiol. 35:3071-3077.[Abstract]
  4. Lauretti, L., M. L. Riccio, A. Mazzariol, G. Cornaglia, G. Amicosante, R. Fontana, and G. M. Rossolini. 1999. Cloning and characterization of blaVIM, a new integron-borne metallo-ß-lactamase gene from a Pseudomonas aeruginosa clinical isolate. Antimicrob. Agents Chemother. 43:1584-1590.[Abstract/Free Full Text]
  5. Lévesque, C., L. Piché, C. Larose, and P. H. Roy. 1995. PCR mapping of integrons reveals several novel combinations of resistance genes. Antimicrob. Agents Chemother. 39:185-191.[Abstract]
  6. Nordmann, P., and L. Poirel. 2002. Emerging carbapenemases in gram negative aerobes. Clin. Microbiol. Infect. 8:321-331.[CrossRef][Medline]
  7. Poirel, L., T. Naas, D. Nicolas, L. Collet, S. Bellais, J.-D. Cavallo, and P. Nordmann. 2000. Characterization of VIM-2, a carbapenem-hydrolyzing metallo-ß-lactamase and its plasmid- and integron-borne gene from a Pseudomonas aeruginosa clinical isolate in France. Antimicrob. Agents Chemother. 44:891-897.[Abstract/Free Full Text]
  8. Rossolini, G. M., M. L. Riccio, G. Cornaglia, L. Pagani, C. Lagatolla, L. Selan, and R. Fontana. 2000. Carbapenem-resistant Pseudomonas aeruginosa with acquired blaVIM metallo-ß-lactamase determinants, Italy. Emerg. Infect. Dis. 6:312-313.[Medline]
  9. Sambrook, J., and D. W. Russell. 2001. Molecular cloning: a laboratory manual, 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
  10. Walsh, T. R., A. Bolmström, A. Qwärnström, and A. Gales. 2002. Evaluation of a new Etest for detecting metallo-ß-lactamases in routine clinical testing. J. Clin. Microbiol. 40:2755-2759.[Abstract/Free Full Text]
  11. Yatsuyanagi, J., S. Saito, S. Harata, N. Suzuki, Y. Ito, K. Amano, and K. Enomoto. 2004. Class 1 integron containing metallo-ß-lactamase gene blaVIM-2 in Pseudomonas aeruginosa clinical strains isolated in Japan. Antimicrob. Agents Chemother. 48:626-628.[Abstract/Free Full Text]
T. Conceição
A. Brízio
A. Duarte*
Laboratório de Microbiologia
Faculty of Pharmacy
Av. Forças Armadas 1649-019
Lisbon, Portugal

R. Barros
Hospital Dona EstefÂnia
Lisbon, Portugal

* Phone: 351 21 7946440,Fax: 351 21 7986055,E-mail: aduarte{at}ff.ul.pt

Antimicrobial Agents and Chemotherapy, January 2005, p. 476, Vol. 49, No. 1
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.1.476.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:


This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Conceição, T.
Right arrow Articles by Barros, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Conceição, T.
Right arrow Articles by Barros, R.

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Clin. Vaccine Immunol. Clin. Microbiol. Rev.
J. Clin. Microbiol. ALL ASM JOURNALS