Surface disinfection together with cleaning practises and aseptic procedures are established measures for effective prevention of infectious diseases reducing infection risks. In this study, we evaluated the bactericidal effectiveness in vitro of the electro-medical device Sani System, an over-heated saturated dry-steam disinfection system, against predetermined bacterial load dried on inert surfaces. In particular we have tested different materials, representative of operating rooms furnishing and walls commonly used in healthcare setting, with Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii. As a result of treatment, the mean bacterial load on all the test surfaces was reduced by a factor of 102 in the contact plate experimental study and by a factor of 10 in the total bioburden experimental study. The Sani System Polti proved to be efficacious in killing 100% of the bacteria in all experimental conditions, therefore it could help to reduce the risk of spreading nosocomial infections in healthcare facilities.