In order to estimate the clinical efficacy of a povidone-iodine oral antiseptic (PVP-I) on oral bacterial infectious diseases, we studied the effect of oral organic matter on the in vitro killing activity of PVP-I. In addition, we compared the in vitro short-time killing activity of PVP-I with those of other oral antiseptics using mouth-washing and gargling samples collected from healthy volunteers. When any of the mouth-washing and gargling samples was used, the standard (0.23-0.47%) or lower concentrations of PVP-I killed methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa, including multidrug-resistant strains, within 15-60 s in the presence of oral organic matter. 0.02% benzethonium chloride (BEC) and 0.002% chlorhexidine gluconate (CHG) did not show effects against MRSA and P. aeruginosa (including multidrug-resistant strains) in mouth-washing and gargling samples even after 60 s. The above-mentioned results show that the in vitro killing activity of the standard concentration of PVP-I was hardly affected by the oral organic matter and that a mouth-washing or gargling solution containing PVP-I has a stronger bactericidal activity than BEC and CHG. Although mouth-washing and gargling samples were obtained from healthy individuals in this study, PVP-I may be used for protection against infections in patients with various diseases, if proper concentrations and usage are encouraged.