The aims of this study are as follows: (1) to investigate the level and probability of bacterial contamination on scrub suits over time for medical personnel working inside and/or outside the operating room (OR) area; (2) to discuss the protective role of cover gowns against bacterial contamination; and (3) to consider the necessity of changing into clean suits whenever entering the OR in terms of preventing periprosthetic joint infection (PJI) in total joint replacement (TJR). The bacterial colony count was examined on the chest area of the scrub suits worn during various daily clinical practices. The genus/species of the contaminants were identified by matrix-assisted laser deposition ionization-time of flight mass spectrometry (MALDI-TOF MS). The scrub suits worn outside the OR area showed a greater level of bacterial contamination than those worn inside the OR area, although the colony counts tended to increase over time both in and out of the area. The probability of contamination involving coagulase-negative staphylococci (CNS) (known as the primary cause of PJI) did not increase significantly in the scrubs worn inside the OR area, but did increase significantly after a long-time departure from the OR area. Our results suggest that wearing scrub suits outside and/or the long duration of wearing the same suits can significantly increase the level and probability of any bacterial contamination (including CNS) on scrub suits. We also found that wearing a cover gown over scrub suits outside the OR area may have only a limited protective role against bacterial contamination.