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Journal of Clinical Microbiology, February 2000, p. 542-546, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Detection of Extended-Spectrum beta -Lactamases in Clinical Isolates of Enterobacter cloacae and Enterobacter aerogenes

Eva Tzelepi,1 Panagiota Giakkoupi,1 Danai Sofianou,2 Veneta Loukova,1 Anastassia Kemeroglou,2 and Athanassios Tsakris3,*

Department of Bacteriology, Hellenic Pasteur Institute, 115 21 Athens,1 Department of Microbiology, Hippokration General Hospital, 54642 Thessaloniki,2 and Department of Microbiology, Medical School, Aristotle University of Thessaloniki, 54006 Thessaloniki,3 Greece

Received 12 July 1999/Returned for modification 30 September 1999/Accepted 28 October 1999

The aim of the present study was to investigate the frequency of extended-spectrum beta -lactamases (ESBLs) in a consecutive collection of clinical isolates of Enterobacter spp. The abilities of various screening methods to detect ESBLs in enterobacters were simultaneously tested. Among the 68 consecutive isolates (56 Enterobacter cloacae and 12 Enterobacter aerogenes isolates) that were analyzed for beta -lactamase content, 21 (25 and 58%, respectively) possessed transferable ESBLs with pIs of 8.2 and phenotypic characteristics of SHV-type enzymes, 8 (14.3%) of the E. cloacae isolates produced a previously nondescribed, clavulanate-susceptible ESBL that exhibited a pI of 6.9 and that conferred a ceftazidime resistance phenotype on Escherichia coli transconjugants, and 2 E. cloacae isolates produced both of these enzymes. Among the total of 31 isolates that were considered ESBL producers, the Vitek ESBL detection test was positive for 2 (6.5%) strains, and the conventional double-disk synergy test (DDST) with amoxicillin-clavulanate and with expanded-spectrum cephalosporins and aztreonam was positive for 5 (16%) strains. Modifications of the DDST consisting of closer application of the disks (at 20 instead of 30 mm), the use of cefepime, and the use of both modifications increased the sensitivity of this test to 71, 61, and 90%, respectively. Of the 37 isolates for which isoelectric focusing failed to determine ESBLs, the Vitek test was false positive for 1 isolate and the various forms of DDSTs were false-positive for 3 isolates.

* Corresponding author. Mailing address: Department of Microbiology, Medical School, Aristotle University of Thessaloniki, 54006-Thessaloniki, Greece. Phone: 30 31 99 90 91. Fax: 30 31 99 91 49. E-mail: atsakris{at}med.auth.gr.

Journal of Clinical Microbiology, February 2000, p. 542-546, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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