ABSTRACTWe characterized five carbapenemase-producingEnterobacterales(CPE) isolates from two health care institutions in Lima, Peru. The isolates were identified asKlebsiella pneumoniae(n= 3),Citrobacter portucalensis(n= 1), andEscherichia coli(n= 1). All were identified asblaOXA-48-like gene carriers using conventional PCR. Whole-genome sequencing found the presence of theblaOXA-181gene as the only carbapenemase gene in all isolates. Genes associated with resistance to aminoglycosides, quinolones, amphenicols, fosfomycins, macrolides, tetracyclines, sulfonamides, and trimethoprim were also found. The plasmid incompatibility group IncX3 was identified in all genomes in a truncated Tn6361transposon flanked by ΔIS26insertion sequences. TheqnrS1gene was also found downstream ofblaOXA-181, conferring fluoroquinolone resistance to all isolates. CPE isolates harboringblaOXA-like genes are an increasing public health problem in health care settings worldwide. The IncX3 plasmid is involved in the worldwide dissemination ofblaOXA-181, and its presence in these CPE isolates suggests the wide dissemination ofblaOXA-181in Peru.IMPORTANCEReports of carbapenemase-producingEnterobacterales(CPE) isolates are increasing worldwide. Accurate detection of the β-lactamase OXA-181 (a variant of OXA-48) is important to initiate therapy and preventive measures in the clinic. OXA-181 has been described in CPE isolates in many countries, often associated with nosocomial outbreaks. However, the circulation of this carbapenemase has yet to be reported in Peru. Here, we report the detection of five multidrug-resistant CPE clinical isolates harboringblaOXA-181in the IncX3-type plasmid, a potential driver of dissemination in Peru.