Genetic Structure Associated with blaOXA-18, Encoding a Clavulanic Acid-Inhibited Extended-Spectrum Oxacillinase

The genetic environment of the bla(OXA-18) gene encoding a peculiar clavulanic acid-inhibitable Ambler class D extended-spectrum beta-lactamase was determined from the prototype OXA-18-producing Pseudomonas aeruginosa MUS clinical isolate. An 8.2-kb genomic DNA fragment containing blaOXA-18 was cloned from P. aeruginosa MUS. Although most oxacillinases are located in integrons, blaOXA-18 lacked gene cassette-specific features. It was bracketed by two duplicated sequences containing ISCR19, a novel insertion sequence of the ISCR family of mobile elements; Delta intI1, a truncated integrase gene; and a truncated Delta aac6'-Ib gene cassette. It is likely that ISCR19 was at the origin of the bla(OXA-18) gene mobilization by a rolling-circle transposition event followed by homologous recombination. Furthermore, analysis of the cloned genomic DNA fragment revealed the presence of the integron-containing bla(OXA-20) gene. Concomitantly, three P. aeruginosa clinical isolates, displaying a synergy image as determined by double-disk diffusion tests on cloxacillin-containing plates, were isolated from three patients hospitalized in different wards over a 9-month period at the Saint-Luc University hospital (Brussels, Belgium). These isolates were positive by PCR for bla(OXA-18) and bla(OXA-20) genes, genetically related to P. aeruginosa MUS as determined by pulsed-field gel electrophoresis, and carried the same bla(OXA-18)/bla(OXA-20)-associated genetic structures. This report characterized the genetic elements likely at the origin of blaOXA-18 gene mobilization in P. aeruginosa and suggests the spread of oxacillin-type extended-spectrum beta-lactamases in P. aeruginosa at the Saint-Luc University hospital of Brussels, Belgium.