Background: Recently, the probable involvement of surfaces from the hospital environment as a disseminating source of resistant bacteria has been highlighted. The aim of the study was to compare resistant microorganisms isolated from inanimate surfaces, equipments and patient blood culture samples in an Intensive Care Unit from Belo Horizonte, Brazil. Methods: A cross-sectional study was performed from July to October 2009. Data sources were microbiologic samples from environment and patient blood culture. Duplicate samples were obtained by swabs from up to seven different touch sites around two different patients in four different days. Jointly with the environmental samples, bacterial isolates from an adult ICU patients’ routine blood cultures were obtained from hospital laboratory. The samples were identified, tested for sensitivity and compared by rep-PCR test to verify similarity. Results: Difference among the averages of Colony Forming Units was found within the environment samples (p < 0.004). In the environment were identified antibiotic resistant microorganisms such as Vancomycin resistant Enterococcus faecalis, imipenem and ciprofloxacin Pseudomonas aeruginosa and multidrug-resistant Acinetobacter baumannii. Similarities (60% -80%) were established among environmental and blood culture samples. Conclusion: The environmental sampling showed different averages of contamination of the surfaces and equipment. The similarity among the bacterial isolates of patients’ blood cultures and environmental samples reinforces the hypothesis of the horizontal transference of pathogens.