The mechanisms at the origin of heterogeneous carbapenem resistance levels observed among Acinetobacter baumannii isolates collected in 2005 in a large University Hospital of Rome, Italy, were investigated. These isolates were related and possessed similar plasmids carrying the carbapenem-hydrolyzing oxacillinase gene bla(OXA-58) but showed variable levels of resistance to carbapenems. Analysis of sequences surrounding the bla(OXA-58) gene showed genetic variability, with the presence in several isolates of multiple copies of the bla(OXA-58) gene; this extra copy number was likely related to an IS26-mediated transposition or recombination process.