Background: Hepatitis C virus infection is a significant problem among patients undergoing maintenance hemodialysis. We conducted a prospective multi-center study to evaluate the effect of isolation policy on the spread of HCV infection. Methods:Our study included 114 HCV seronegative patients from four hemodialysis units classified into two groups; the first group included (46) patients undergoing regular hemodialysis in two of these units following strict isolation of the HCV seropositive patients, and the second group included (68) patients undergoing regular hemodialysis in the other two units not following these strict precautions of the HCV positive patients isolation, all these patients were traced over a period of 36 months in these units to determine the incidence of seroconversion.Results: During the follow up period, the incidence of HCV infection in the first group was (14.8%) and it was (42.9%) in the second group.Conclusions: In this study the incidence of HCV in HD patients decreased by the use of isolation policy for HCV infected patients. Additional studies may help to clarify the role of isolation in conjunction with application of universal precautions in reducing HCV transmission.