89 patients undergoing spine surgery were randomized to two different protocols of povidone-iodine application: one group received application just prior to incision, while the other group received application prior to the operating surgeon's hands being scrubbed. The purpose of this study was to evaluate and compare the two protocols in terms of incidence of positive cultures isolated from the surgical site, and the incidence of surgical site infection. The results demonstrated a significantly lower incidence of positive cultures in patients who received povidone-iodine application prior to the surgeon's hands being scrubbed, although there was no significant difference between groups in overall development of surgical site infection.