Abstract: Objective To determine the related risk factors influencing incidence of human immunodeficiency virus (HIV) infection among drug users(DUs) in Yunnan Province and to provide scientific reference for the prevention and control of acquired immune deficiency syndrome(AIDS). Methods HIV-1 antibody positive serum samples of DUs from drug abuse sentinel sites in Yunnan Province during 2013 to 2015 were collected and tested by BED-capture enzyme immunoassay (BED-CEIA). Questionnaires including demographic and behavioral characteristics were performed among DUs to analyze HIV infection risk factors. Results In 31 503 samples, 3 424 were HIV-1 antibody positive(including 2 793 previous infections) and the positive rate was 10.9%(3 424/31 503).Male and female DUs accounted for 87.3% (2 989/3 424) and 12.7% (435/3 424) respectively. Of total 598 new reported HIV cases, 101 new infections were detected by BED-CEIA, among which 90 (89.1%) were male and 11(10.9%) were female with an average age of 31.54 years old (16-54). HIV-1 incident rate among DUs was 0.53% from 2013 to 2015. The result of multivariate Logistic regression analyses showed that new infection risk among participants with the foreign nationality(Myanmar), non-Han nationality and injection drug use was significantly higher than that among participants with registered permanent residence in or not in Yunnan Province,Han nationality and oral drug use (all P<0.05). Conclusions More attention should be paid to DUs with foreign nationality(Myanmar),non-Han nationality and injection drug use. Comprehensive intervention measures including community maintenance treatment, peer education and HIV testing and counseling should be taken to reduce HIV-1 new infection rate.