The natural rubber latex (NRL) film taken from medical surgical gloves was surface-modified with a dielectric barrier discharge (DBD) plasma treatment under an air environment. The results showed that surface hydrophilicity of the NRL film increased after the plasma treatment due to the presence of oxygen-containing polar groups on the plasma-treated surface. An increase in plasma treatment time increased the surface roughness of the NRL film, and eventually decreased the mechanical properties. From the obtained results, the optimum plasma treatment time of 20 s was chosen. After immersion in a chitosan solution, the amount of chitosan deposited on the plasma-treated NRL film increased with increasing chitosan concentrations. The chitosan coating smoothed the surface of the plasma-treated NRL film and also improved the mechanical properties. The highest antibacterial activities of the chitosan-coated DBD plasma-treated NRL film against both Staphylococcus aureus and Escherichia coli were achieved when a 2 %(w/v) chitosan solution was used for the coating.