This study examined the influence of variables in a finishing process for making cotton fabric with regenerable antibacterial properties against Staphylococcus aureus (S. aureus). 5,5-dimethylhydantoin (DMH) was coated onto cotton fabric by a pad-dry-plasma-cure method. Sodium hypochlorite was used for chlorinating the DMH coated fabric in order to introduce antibacterial properties. An orthogonal array testing strategy (OATS) was used in the finishing process for finding the optimum treatment conditions. After finishing, UV-Visible spectroscopy, Scanning Electron Microscopy (SEM), and Fourier Transform Infrared Spectroscopy (FTIR) were employed to characterise the properties of the treated cotton fabric, including the concentration of chlorine, morphological properties, and functional groups. The results show that cotton fabric coated with DMH followed by plasma treatment and chlorination can inhibit S. aureus and that the antibacterial property is regenerable.