Spread of carbapenem-resistant Acinetobacter baumannii carrying a plasmid with two genes encoding OXA-72 carbapenemase in Lithuanian hospitals

OBJECTIVES: To study the molecular epidemiology of Acinetobacter baumannii isolates from Lithuanian hospitals with an emphasis on the characterization of plasmids and antibiotic-resistance genes and their relationship with European clones (ECs) I and II. METHODS: PFGE, PCR analysis of ECs and resistance genes, plasmid replicon typing, DNA transformation and sequencing were employed to characterize A. baumannii. RESULTS: Of the 444 isolates studied, 230 (52%) and 202 (45%) belonged to ECI and ECII clones, respectively, and showed clone-specific resistance gene profiles. Five plasmids from 6 to 100 kb in size in different combinations (one to four plasmids) were found in A. baumannii isolates, the combination of 9 + 70 kb plasmids in ECI isolates (60%, 137/230) and an 11 kb plasmid in ECII isolates (52%, 106/202) being the most frequent. GR2 and GR6 replicon groups, alone or in combination, were found, with a prevalence of GR2 + GR6 in ECI isolates of 90% (206/230) and a prevalence of GR2 in ECII isolates of 56% (113/202). The vast majority (95%, 165/174) of carbapenem-resistant A. baumannii ECII isolates carried a novel GR2-type plasmid of 11 kb, designated pAB120, which had two copies of a blaOXA-72 gene, flanked by XerC/XerD-like sites and conferred resistance to carbapenems when introduced into a carbapenem-susceptible A. baumannii strain. CONCLUSIONS: The spread of carbapenem-resistant A. baumannii in Lithuanian hospitals is strongly associated with strains belonging to ECII and carrying a GR2 plasmid encoding two blaOXA-72 genes. The genetic environment of pAB120 supports the role of site-specific recombination associated with the acquisition of carbapenem-hydrolysing class D β-lactamases.