Objective The study was conducted to determine the prevalence of Staphylococcus aureus and the resistance to commonly used antibiotics in the intensive care unit (ICU), provide the evidence for the prevention, control and clinical treatment. Methods Retrospective analysis was performed to evaluate the epidemiological characteristics and drug resistance of Staphylococcus aureus isolated in the ICU of a hospital from 2005 to 2007. Results A total of 378 strains of Staphylococcus aureus were isolated in the three years, accounting for 7.5% (83/1113), 8.9% (122/1366) and 16.9% (173/1023) of all strains isolated in ICU. In vitro susceptibility tests, the sensitive rate of Staphylococcus aureus to vancomycin and teicoplanin were 100.0%, the resistant rate to nitrofurantoin was 4.8%-9.1%, and the resistant rate to cotrimoxazole from 33.9% to 30.3%. The resistant rates to ampicillin / sulbactam, oxacillin, cefazolin, ciprofloxacin, levofloxacin, gentamicin, clindamycin, erythromycin, tetracycline and rifampin were higher than 50.0%, the resistance to penicillin was up to 97.6%-100.0%. Conclusion The strains of Staphylococcus aureus isolated in ICU increased year by year, but no significant increase in the proportion of MRSA was observed; no MRSA was found to be resistant to vancomycin and teicoplanin. The resistant rates of Staphylococcus aureus to nitrofurantoin and cotrimoxazole were slightly lower, but the resistant rates to other antibiotics were higher. Therefore, attention shall be paid to etiological examination and drug susceptibility test for the effective prevention and control of the prevalence of Staphylococcus aureus and the rational use of antibiotics.