Purpose: To determine the effect of washing the biopsy needle with povidone-iodine solution on infection rates after transrectal ultrasonography-guided prostate biopsy. Patients and Methods: 180 patients with transrectal prostate biopsy were included. Infection was excluded with midstream urine culture before biopsy, and patients were divided into two groups. In group 1 (n = 84), the needle was washed with povidone-iodine after each material reception during biopsy, and in group 2 (n = 96), it was not. At day 3 and 2 weeks after the biopsy, patients were checked. On day 3, midstream urine was cultured and patients were checked for fever, hematuria, rectal hemorrhagia, hematospermia and urinary retention. Also, the effect of rectal preparation on infection rate was evaluated. Results: There was no significant difference between the patients’ ages, PSA levels and prostate volumes. In total, 11 patients (6.1%; 4 from group 1 and 7 from group 2) developed infectious complications. There was no statistically significant difference between the groups. Fever, asymptomatic bacteriuria and urinary system infection rates were also not statistically different between the groups. The most frequent complication was hematuria with 83 cases (46.1%), but they did not need hospitalization and no patient had acute urinary obstruction. Conclusions: Washing the biopsy needle with povidone-iodine did not have an effect on the infection rates. Antibiotic prophylaxis and cleaning the biopsy equipment is adequate for low infection rates.