The use of radiopharmaceuticals for the diagnosis of infection is increasing due to their ability to distinguish between septic and aseptic inflammation. The aim of this study was to analyze the intracellular accumulation of technetium-99m-ciprofloxacin in strains of Staphylococcus aureus and Pseudomonas aeruginosa harboring an overexpression of NorA and MexAB-OprM, respectively.