To evaluate the usefulness of the waterless hand washing method for surgical anti-sepsis, we conducted a microbial sampling study, comparing it to a conventional surgical scrub method. A total of 18 operating-room nurses were undertaking the following three-hand washing protocols : scrubbing with blushes using 4w/v% chlorhexidine gluconate (CHG) and also rubbing with CHG, followed by application of a 0.2w/v% CHG with ethanol (HS) preparation. (conventional method); rubbing with CHG and application of HS (two-stage surgical scrub method; TSS); rubbing with anti-septic soap and application of HS (waterless method; WL). Microbial sampling was conducted after hand washing using the glove juice method. No statistically significant differences in bacterial numbers were found among these three methods. The number of bacterially positive subjects was significantly higher in the conventional method than the TSS method. These results indicate that there are adverse effects of blush-scrubbing, as the detected bacteria were related to normal skin flora. As such, the WL method for hand anti-sepsis appears to be equivalent to the conventional surgical scrubbing method in terms of microbial detection. The WL method should therefore be introduced as a standard hand anti-sepsis method at the time of surgery because it is cost-effective as well as time-efficient.