We assessed the impact of antibiotic administration prior to sample collection on the bacterial resistance rates from patients with nosocomial infection. Every individual susceptibility report was assessed in real time at the bedside of the patient by a team composed of infectious diseases and internal medicine specialists as well as clinical microbiologists for clinical significance and appropriateness of the specimen. The report also stated the kind, source and origin of the infection, history of administration of any antibiotic during the last month prior to sample collection. To evaluate the impact of previous antibiotic administration, resistance rates were calculated separately among the group of patients with and without history of antibiotic treatment. A crude univariate analysis was performed to assess the significance of the differences between groups for every species-antibiotic pair. Patients who had received ciprofloxacin showed significantly higher rates of Escherichia coli resistant to ciprofloxacin, broad-spectrum cephalosporins and gentamicin. A higher rate of methicillin-resistant Staphylococcus aureus was observed in patients who were given gentamicin. A stratified analysis showed that the previous antibiotic administration continued to be a risk factor for increased resistance rates regardless of the hospital ward or the source of the infection. This study demonstrates the influence of previous antibiotic administration on bacterial resistance rates although this fact is barely taken into account by the laboratory when constructing the cumulative susceptibility data. Real time clinical validation of the individual susceptibility reports, performed by a multidisciplinary team prior to the data entering, might be a suitable approach to get more reliable susceptibility rates to guide the rational selection of antimicrobial empirical therapy in patients with hospital-acquired infections who have been given antimicrobial treatment prior to specimen collection.