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Antimicrobial Agents and Chemotherapy, July 1999, p. 1584-1590, Vol. 43, No. 7
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Cloning and Characterization of blaVIM, a New Integron-Borne Metallo-beta -Lactamase Gene from a Pseudomonas aeruginosa Clinical Isolate

Laura Lauretti,1 Maria Letizia Riccio,1 Annarita Mazzariol,2 Giuseppe Cornaglia,2 Gianfranco Amicosante,3 Roberta Fontana,2 and Gian Maria Rossolini1,*

Dipartimento di Biologia Molecolare, Sezione di Microbiologia, Università di Siena, 53100-Siena,1 Istituto di Microbiologia, Università di Verona, 37134-Verona,2 and Dipartimento di Scienze e Tecnologie Biomediche e Biometria, Università dell'Aquila, 67100-L'Aquila,3 Italy

Received 8 October 1998/Returned for modification 30 January 1999/Accepted 17 April 1999

Production of a metallo-beta -lactamase activity was detected in a carbapenem-resistant Pseudomonas aeruginosa clinical isolate (isolate VR-143/97) from an Italian inpatient at the Verona University Hospital (northern Italy). The metallo-beta -lactamase determinant was isolated from a genomic library of VR-143/97, constructed in an Escherichia coli plasmid vector, by screening for clones with reduced susceptibility to imipenem. Sequencing of the cloned gene revealed that it encoded a new class B beta -lactamase that was named VIM-1. At the sequence level VIM-1 was rather divergent from the other class B enzymes (16.4 to 38.7% identity), overall being more similar to members of subclass B1 including the beta -lactamase II of Bacillus cereus (Bc-II), the Bacteroides fragilis CcrA, the Chryseobacterium meningosepticum BlaB, and the cassette-encoded IMP-1 enzymes. Among these, VIM-1 showed the highest degree of similarity to Bc-II. Similarly to blaIMP, blaVIM was also found to be carried on a gene cassette inserted into a class 1 integron. The blaVIM-containing integron was located on the chromosome of P. aeruginosa VR-143/97, and the metallo-beta -lactamase-encoding determinant was not transferable to E. coli by conjugation. Expression of the integron-borne blaVIM gene in E. coli resulted in a significant decrease in susceptibility to a broad array of beta -lactams (ampicillin, carbenicillin, piperacillin, mezlocillin, cefotaxime, cefoxitin, ceftazidime, cefoperazone, cefepime, and carbapenems), revealing a very broad substrate specificity of the VIM-1 enzyme.

* Corresponding author. Mailing address: Dipartimento di Biologia Molecolare, Sez. di Microbiologia, Università di Siena, Via Laterina, 8, 53100-Siena, Italy. Phone: 39 0577 233327. Fax: 39 0577 233325. E-mail: rossolini{at}unisi.it.

Antimicrobial Agents and Chemotherapy, July 1999, p. 1584-1590, Vol. 43, No. 7
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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