Objective To analyze the infection and risk factors of hepatitis C virus (HCV) among drug users. Methods Drug users from drug rehabilitation center, methadone clinic, and community of Jinghong City, Xishuangbanna Autonomous Prefecture, Yunnan Province were recruited by researchers from January to July 2021 according to the inclusion and exclusion criteria. A structured questionnaire was adopted, 3 mL serum specimens were taken from the respondents, HCV serological screening test was performed with enzyme-linked immunosorbent assay (ELISA). HCV infection and its risk factors among drug users in Jinghong City, Xishuangbanna Autonomous Prefecture, Yunnan Province were analyzed. Results A total of 404 drug users were investigated, 26 (6.44%) were HCV antibody positive. Intravenous drug injection (OR=4.236, 95%CI: 1.644-10.913, P=0.003), female (OR=0.135, 95%CI: 0.019-0.987, P=0.048), as well as high school and above education (OR=6.745, 95%CI: 1.113-40.890, P=0.038) drug users had a higher risk of HCV infection, differences were all statistica-lly significant (all P < 0.05). Conclusion Intravenous drug injection is a high risk factor for HCV infection among drug users in Jinghong City, Xishuangbanna Autonomous Prefecture, Yunnan Province, the age, educational level and gender of drug users are also related to HCV infection.