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Antimicrobial Agents and Chemotherapy, December 2003, p. 3739-3742, Vol. 47, No. 12
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.12.3739-3742.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

In Vivo Selection of a Chromosomally Encoded ß-Lactamase Variant Conferring Ceftazidime Resistance in Klebsiella oxytoca

Hedi Mammeri, Laurent Poirel, and Patrice Nordmann*

Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, Université Paris XI, 94275 Le Kremlin-Bicêtre, France

Received 27 May 2003/ Returned for modification 16 July 2003/ Accepted 22 September 2003

Klebsiella oxytoca clinical isolate A was recovered from the urine of a 55-year-old man with prostatic and urinary tract infections. This isolate displayed a ß-lactam resistance phenotype consistent with overproduction of a chromosomally encoded class A ß-lactamase and had decreased susceptibilities to all ß-lactams except ceftazidime, cephamycins, and carbapenems. Four weeks after treatment with an antibiotic regimen that included ceftazidime, K. oxytoca isolate B, which had a high level of resistance to ceftazidime, was isolated from the urine of the same patient. Isoelectric focusing analysis of the culture extracts of these isolates gave a pI of 5.4 for both isolates. Cloning experiments with the PCR products of the blaOXY gene resulted in two Escherichia coli DH10B recombinant clones with resistance phenotypes mirroring those of the parental isolates. Sequencing analysis revealed that the blaOXY-2-5 gene from K. oxytoca B had a single nucleotide substitution compared to the sequence of the blaOXY-2 gene from K. oxytoca A, leading to a proline-to-serine substitution at position 167, according to the numbering of Ambler. Biochemical analysis of purified OXY-2-5 showed that it had the ability to hydrolyze ceftazidime. This is the first report of in vivo selection of a K. oxytoca isolate that produced a chromosomally encoded ß-lactamase conferring resistance to ceftazidime.

* Corresponding author. Mailing address: Service de Bactériologie-Virologie, Hôpital de Bicêtre, 78 rue du Général Leclerc, 94275 Le Kremlin-Bicêtre Cedex, France. Phone: 33-1-45-21-36-32. Fax: 33-1-45-21-63-40. E-mail: nordmann.patrice{at}bct.ap-hop-paris.fr.

Antimicrobial Agents and Chemotherapy, December 2003, p. 3739-3742, Vol. 47, No. 12
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.12.3739-3742.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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