To evaluate the incidence and risk factors for human immunodeficiency virus (HIV) seroconversion of HIV-negative partners among HIV-discordant couples in Liuzhou, China, 1854 eligible HIV-serodiscordant couples were retrospectively identified through the HIV epidemiology and follow-up database from January 1, 1996 to June 30, 2013. Cox proportional-hazards model was used to examine risk factors related to HIV seroconversion of negative partners. Finally, 125 HIV seroconversion occurred over 4963.5 person-years, resulting in an overall HIV incidence of 2.52/100 person-years. HIV-positive partners with the last CD4 counts of 350 cells/ul or more were significantly protected against HIV seroconversion compared with those CD4 counts of less than 200 cells/ul (aHR = 0.46, 95% CI: 0.27–0.81, P < 0.01). Men with HIV-positive wives (aHR: 1.96, 95% CI: 1.27–3.02, P < 0.01), HIV-positive partners who did not receive ART before their HIV-negative partners' seroconversion (aHR: 2.22, 95% CI, 1.41–3.51, P < 0.01) and patients reported intermittent condom use (aHR: 7.60, 95% CI, 4.37–13.21, P < 0.01) were associated with increased risk of HIV seroconversion. HIV-negative partners remain high risk of HIV infection in Liuzhou city. Comprehensive package of HIV prevention services should contribute to reduction in HIV transmission of discordant couples. J. Med. Virol. 87:553–556, 2015. © 2015 Wiley Periodicals, Inc.