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Antimicrobial Agents and Chemotherapy, November 2005, p. 4716-4720, Vol. 49, No. 11
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.11.4716-4720.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular Analysis of the Simultaneous Production of Two SHV-Type Extended-Spectrum Beta-Lactamases in a Clinical Isolate of Enterobacter cloacae by Using Single-Nucleotide Polymorphism Genotyping

Dóra Szabó,1,2 Melissa A. Melan,3 Andrea M. Hujer,4 Robert A. Bonomo,4 Kristine M. Hujer,4 Christopher R. Bethel,4 Katalin Kristóf,1 and David L. Paterson2*

Institute of Medical Microbiology, Semmelweis University, Budapest, Hungary,1 Division of Infectious Diseases, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania 15213,2 Department of Pathology, Division of Molecular Diagnostics, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania 15213,3 Research Service, Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, Ohio 441064

Received 28 March 2005/ Returned for modification 12 June 2005/ Accepted 25 August 2005

Bacteria that simultaneously produce multiple extended-spectrum beta-lactamases are frequently isolated. We report an Enterobacter cloacae isolate, ES24, producing four different beta-lactamases (AmpC type beta-lactamase, TEM-1, SHV-7, and a novel extended-spectrum beta-lactamase, SHV-30). Direct sequencing of blaSHV gene products gave a "double peak" at position 703, suggesting the presence of more than one allele. Using fluorescence resonance energy transfer real-time PCR to detect single-nucleotide polymorphisms, we were able to distinguish two different blaSHV genes in a single isolate. This may prove to be a useful technique in surveys of beta-lactamase production in contemporary clinical isolates.

* Corresponding author. Mailing address: Division of Infectious Diseases, University of Pittsburgh Medical Center, Suite 3A Falk Medical Building, 3601 5th Avenue, Pittsburgh, PA 15213. Phone: (412) 648-6478. Fax: (412) 648-6399. E-mail: patersond{at}dom.pitt.edu.

Antimicrobial Agents and Chemotherapy, November 2005, p. 4716-4720, Vol. 49, No. 11
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.11.4716-4720.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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