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Antimicrobial Agents and Chemotherapy, May 1999, p. 1206-1210, Vol. 43, No. 5
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Carbapenem Resistance in Escherichia coli Associated with Plasmid-Determined CMY-4 beta -Lactamase Production and Loss of an Outer Membrane Protein

Paul D. Stapleton,* Kevin P. Shannon, and Gary L. French

Department of Microbiology, UMDS, St. Thomas' Hospital, London, United Kingdom

Received 22 July 1998/Returned for modification 12 November 1998/Accepted 10 March 1999

Three cefoxitin-resistant Escherichia coli isolates from stool specimens of a patient with leukemia were either resistant, intermediate, or sensitive to imipenem. Conjugation experiments showed that cefoxitin resistance, but not imipenem resistance, was transferable. All isolates were shown by isoelectric focusing to produce two beta -lactamases with isoelectric points of 5.4 (TEM-1, confirmed by sequencing of a PCR product) and >8.5 (consistent with a class C beta -lactamase). The gene coding for the unknown beta -lactamase was cloned and sequenced and revealed an enzyme which had 99.9% sequence identity with the plasmid-determined class C beta -lactamase CMY-2. The cloned beta -lactamase gene differed from blaCMY-2 at one nucleotide position that resulted in an amino acid change, tryptophan to arginine at position 221. We propose that this enzyme be designated CMY-4. Both the imipenem-resistant and -intermediate isolates lacked a 38-kDa outer membrane protein (OMP) that was present in the imipenem-sensitive isolate. The lack of an OMP alone did not explain the difference in carbapenem susceptibilities observed. However, measurement of beta -lactamase activities (including measurements under conditions where TEM-1 beta -lactamase was inhibited) indicated that the imipenem-intermediate isolate expressed six- to eightfold less beta -lactamase than did the other isolates. This study illustrates that carbapenem resistance in E. coli can arise from high-level expression of plasmid-mediated class C beta -lactamase combined with an OMP deficiency. Furthermore, in the presence of an OMP deficiency, the level of expression of a plasmid-mediated class C beta -lactamase is an important factor in determining whether E. coli isolates are fully resistant to carbapenems.

* Corresponding author. Mailing address: Department of Biology, Darwin Bldg., University College London, Gower St., London WC1E 6BT, United Kingdom. Phone: 44 171 504 2934. Fax: 44 171 380 7098. E-mail: p.stapleton{at}ucl.ac.uk.

Antimicrobial Agents and Chemotherapy, May 1999, p. 1206-1210, Vol. 43, No. 5
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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