To establish a convenient method for decontaminating bioclean rooms, the effect of ozone at 80 mg/m3 for 72 h was compared with formaldehyde vaporization at an initial concentration of 150 mg/m3 with a gradual decrease to 20 mg/m3 during 72 h. Ozone was found to be inferior to formaldehyde in activity. When the bioclean room was decontaminated twice with ozone, the mean colony count per 10 cm2 was decreased to about the same level as when formaldehyde was used. Ozone had a strong caustic effect upon rubber materials. Despite these disadvantages, ozone decontamination was demonstrated to be superior to formaldehyde vaporization because of convenience, insignificant inhalation of the disinfectant by the hospital staff, and very rapid expulsion of the gas after ventilation. Because the disadvantages of ozone can be easily controlled, this study suggests that ozone decontamination is a promising method for maintaining bioclean rooms.