The nosocomial infections caused by opportunistic pathogen Acinetobacter baumannii was increasing in recent years. It was known that the attachment of A. baumannii on solid surface was a key factor for infection. This study was conducted to evaluate the inhibitory effect of titanium dioxide (TiO2)/Ag thin film on A. baumannii. In this study, TiO2 thin films were firstly prepared by using the modified impregnation method, then TiO2/Ag thin films were prepared by using the spin-coating with 5000 ppm Ag+. The TiO2/Ag thin film was analyzed by field-emission scanning electron micrographs and X-ray diffractometer. The nano-size and Anatase crystal structure were confirmed. Twenty clinical A. baumannii isolates were examined for the bactericidal effect of TiO2/Ag thin film either in the dark or under visible light activation for 20 min. The inhibition efficiencies of TiO2/Ag thin film under visible light on these 20 isolates ranged from 41% to 90%. The effects of TiO2/Ag thin film in the dark were slightly lower than visible light-activated group. Our results suggested that coating of TiO2/Ag on solid surface in hospital environment might be helpful for the prevention of nosocomial A. baumannii infection.