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Journal of Clinical Microbiology, December 2004, p. 5715-5721, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5715-5721.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Phenotypic and Molecular Detection of CTX-M-ß-Lactamases Produced by Escherichia coli and Klebsiella spp.

Johann D. D. Pitout,1 Ashfaque Hossain,2 and Nancy D. Hanson2*

Division of Microbiology, Calgary Laboratory Services, and Department of Pathology & Laboratory Medicine, University of Calgary, Calgary, Alberta, Canada,1 Department of Medical Microbiology and Immunology, Creighton University School of Medicine, Omaha, Nebraska2

Received 10 March 2004/ Returned for modification 12 May 2004/ Accepted 23 August 2004

Organisms producing CTX-M-ß-lactamases are emerging around the world as a source of resistance to oxyiminocephalosporins such as cefotaxime (CTX). However, the laboratory detection of these strains is not well defined. In this study, a molecular detection assay for the identification of CTX-M-ß-lactamase genes was developed and used to investigate the prevalence of these enzymes among clinical isolates of Escherichia coli and Klebsiella species in the Calgary Health Region during 2000 to 2002. In addition, National Committee for Clinical Laboratory Standards (NCCLS) recommendations were evaluated for the ability to detect isolates with CTX-M extended-spectrum ß-lactamases (ESBLs). The PCR assay consisted of four primer sets and demonstrated 100% specificity and sensitivity for detecting different groups of CTX-M-ß-lactamases in control strains producing well-characterized ESBLs. Using these primer sets, 175 clinical strains producing ESBLs were examined for the presence of CTX-M enzymes; 24 (14%) were positive for blaCTX-M-1-like genes, 95 (54%) were positive for blaCTX-M-14-like genes, and the remaining 56 (32%) were negative for blaCTX-M genes. Following the NCCLS recommendations for ESBL testing, all of the control and clinical strains were detected when screened with cefpodoxime and when both cefotaxime and ceftazidime with clavulanate were used as confirmation tests.

* Corresponding author. Mailing address: Department of Medical Microbiology and Immunology, Creighton University School of Medicine, Omaha, NE 68178. Phone: (402) 280-5837. Fax: (402) 280-1875. E-mail: ndhanson{at}creighton.edu.

Journal of Clinical Microbiology, December 2004, p. 5715-5721, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5715-5721.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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