Background: A postoperative infection is one serious complication of an anterior cruciate ligament (ACL) reconstruction. Multiple infection prevention measures have been used before and after surgery, and their clinical efficacy was evaluated in this study. Methods: This study enrolled 54 patients admitted to our hospital from January 2015 to June 2017 who underwent arthroscopic ACL reconstructions with hamstring tendons. One day before the surgery, the bilateral thighs and skin were washed using a potassium permanganate solution before and after bathing, separately. Aseptic surgical films containing iodine and disposable arthroscopic waterproof sterile drapes were used in the surgery. In addition, all the instruments used during the tendon transplantation were soaked and rinsed in a saline solution containing vancomycin. The Lysholm Knee Scoring Scale, International Knee Documentation Committee (IKDC) knee evaluation, and Tegner Activity Scale were administered before the surgery, 3 and 6 months after the surgery, and at the last follow-up. The averages of the scores at different stages were calculated. All the patients received a magnetic resonance imaging examination 3 months after the surgery. Results: The patients were followed-up for 6 to 18 months (average = 10±1.4), and no infections or other complications were found among them. When compared with the preoperative scores, the Lysholm score, IKDC knee evaluation score, and Tegner score were all significantly increased after the operation. Conclusion: With multiple prevention measures adopted, no postoperative infections occurred in the patients, and the knee function recovered well. This suggests the success of the anti-infective therapy following the ACL reconstructions.