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Journal of Antimicrobial Chemotherapy (2003) 52, 1018-1021
© 2003 The British Society for Antimicrobial Chemotherapy

Contribution of ß-lactamase and PBP amino acid substitutions to amoxicillin/clavulanate resistance in ß-lactamase-positive, amoxicillin/clavulanate-resistant Haemophilus influenzae

Vlatka Matic1,*, Bülent Bozdogan1,§, Michael R. Jacobs2, Kimiko Ubukata3 and Peter C. Appelbaum1

1 Department of Pathology, Hershey Medical Center, 500 University Drive, Hershey, PA 17033; 2 Case Western Reserve University, Cleveland, OH, USA; 3 Kitasato University, Tokyo, Japan

Received 9 July 2003; returned 28 August 2003; revised 2 September 2003; accepted 15 September 2003

The roles of ß-lactamase and alterations in penicillin-binding protein in the development of amoxicillin and amoxicillin/clavulanate resistance in two ß-lactamase-positive, amoxicillin/clavulanate-resistant (BLPACR) strains of Haemophilus influenzae were investigated. Seven ß-lactamase-negative, ampicillin-resistant (BLNAR) strains were also studied for comparison of their resistance mechanisms. All strains had been recovered from patients in Japan. The TEM type ß-lactamase of the two BLPACR strains had 100% homology with the amino acid sequences of published TEM-1 ß-lactamase, showing that amoxicillin/clavulanate resistance was not associated with mutations in this ß-lactamase. However, these strains, as well as the seven BLNAR strains, had multiple mutations in ftsI, which encodes penicillin binding protein 3 (PBP3). The transformation of H. influenzae Rd strain with amplified ftsI genes from two BLPACR and two BLNAR strains enabled the selection of amoxicillin/clavulanate-resistant transformants with the same mutations as their parent strains. We concluded that amoxicillin/clavulanate resistance in the two BLPACR strains was due to changes in PBP3. The possibility of the presence of an extended spectrum ß-lactamase was excluded in the BLPACR strains studied.

Keywords: H. influenzae, penicillin-binding proteins, ß-lactamases, mutations

* Present address: PLIVA Inc., Research Institute, Prilaz baruna Filipovica 25, 10000 Zagreb, Croatia

§ Corresponding author. Tel: +1-717-531–3910; Fax: +1-717-531-7953; E-mail: bozdogan-b{at}psu.edu


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