Background:Hepatitis C virus (HCV) infection rates in drug users vary among different regions of China. Drug users who are unaware of their HCV serostatus tend to engage in more risky behaviors.Objectives:This prospective study aimed to assess risk factors of HCV infection in drug users among 11 methadone maintenance treatment (MMT) clinics in Xi’an, China.Patients and Methods:Baseline characteristics and drug use information of patients were collected upon enrollment in the study and anti-HCV tests were performed within one month after the enrollment. Data on daily medication, monthly random urine morphine test results, illicit drug use and MMT retention time were recorded during a 5-year follow-up.Results:Of 10243 patients, 58.0% had positive results for anti-HCV. Injection drug use, longer duration of drug abuse, older age, female gender, unmarried status and unemployment were independent risk factors of HCV infection. Urine test positivity rate was lower (14.8% vs. 16.7%, χ2 = 100.235, P < 0.05), but MMT retention rate was higher (log-rank χ2 = 4.397, P < 0.05) in the anti-HCV positive group than anti-HCV negative one. However, multivariate regression revealed no significant association between anti-HCV serostatus and either MMT retention time or illicit drug use.Conclusions:The major risk factor of HCV infection was injection drug use. The patient’s awareness of his or her HCV status had a minor effect in reduction of illicit drug use and improvement in MMT retention. Therefore, adequate counseling is necessary for drug users in MMT clinics in Xi’an.