High prevalence of OXA-51-type class D β-lactamases among ceftazidime-resistant clinical isolates of Acinetobacter spp.:: co-existence with OXA-58 in multiple centres

Objectives: This study was designed to demonstrate the prevalence of the newly discovered carbapenem-hydrolysing class D enzymes, OXA-51-type and OXA-58, among clinical isolates of Acinetobacter spp. Methods: A total of 72 isolates from six centres were studied. Isolates were screened by PCR with specific primers for bla(OXA-51-type) and bla(OXA-58). PCR products were sequence-analysed. Plasmids were digested with EcoRV and genomic DNAs were digested with PvuII. Hybridization experiments were done with digoxigenin-labelled specific probes. Macro-restriction analysis was done on SmaI-digested genomic DNAs. Results: A total of 56 (77.8%) isolates were positive for bla(OXA-51-type) genes. Sequence analysis of the products from 23 selected isolates revealed the occurrence of multiple alleles in all contributing centres. The bla(OXA-58) gene was detected among 10 isolates from five centres. All were also positive for bla(OXA-51-type) genes. Among the bla(OXA-58)-positive isolates, two from the same centre were positive for a novel OXA-51 allele (OXA-86). Southern hybridization of plasmids and of genomic DNAs suggested that bla(OXA-51-type) genes are located on chromosomes whereas bla(OXA-58) genes are plasmid borne in these 10 isolates. Plasmid profiles and pulsed-field gel electrophoresis patterns indicated the spread of the bla(OXA-58) gene among multiple clones. The bla(OXA-51-type) and bla(OXA-58) co-carrier strains were mostly associated with a pandrug-resistant phenotype. Conclusions: This study indicated that bla(OXA-58)-bearing plasmids are readily spreading among multiple clones of the bla(OXA-51-type)-bearing clinical isolates of Acinetobacter spp. Since these isolates are highly resistant to antibiotics this finding indicates the existence of a significant problem in Turkish hospitals.