Journal of Antimicrobial Chemotherapy (1999) 43, 187-193
© 1999 The British Society for Antimicrobial Chemotherapy
Detection of OXA-4 ß-lactamase in Pseudomonas aeruginosa isolates by genetic methods
a Department of Clinical Pathology, Showa University Fujigaoka Hospital, 1-30 Fujigaoka, Aoba-ku, Yokohama 227-8501; b Department of Biochemistry Sagami Women's University, 2-1-1 Bunkyou, Sagamihara 228-0807; c Department of Biochemistry, Division of Pharmaceutical Science, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142-8555, Japan.
In Pseudomonas aeruginosa, resistance to cefclidin is usually associated with resistance
to another third-generation cephalosporin, ceftazidime. In this study we analysed 22 isolates of P. aeruginosa, collected at Showa University Fujigaoka Hospital between 1992 and
1993, which were resistant to cefclidin but susceptible to ceftazidime. All polymerase chain
reaction (PCR) products amplified by a primer pair covering the full-length gene of OXA-4 (also
OXA-1) precursor ß-lactamase were 0.84 kb in length. The isoelectric points of the
ß-lactamases produced by these isolates were typical of the OXA-4 type of ß-lactamase
(pl 7.5) rather than the OXA-1 type (pI 7.4). All PCR products at 216 bp were amplified by the
primer pair covering the A928
T point mutation, which corresponds to the Asp48Val
amino acid substitution of OXA-1 ß-lactamase to form OXA-4 ß-lactamase. These
single-strand conformation polymorphism (SSCP) patterns are typical of the OXA-4 gene, rather
than the OXA-1 gene, demonstrating that these enzymes can be classified by SSCP analyses
based on the PCR method. Although OXA-4 ß-lactamase is generally plasmid-mediated,
the
chromosomal DNA of these isolates, but not their plasmids, hybridized with the OXA-4 gene
amplified by the PCR method. Based on these results, we suspected that the plasmids encoding
OXA-4 ß-lactamase had been spontaneously cured, or that the gene had been deleted from
the plasmid. The distribution of P. aeruginosa producing OXA-4 ß-lactamase
amongst hospital wards and clinical specimens demonstrated that the OXA-4 enzyme in this
collection period was representative of hospital P. aeruginosa.
* Corresponding author. Tel: +81-45-974-6677; Fax: +81-45-973-1019.
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