Objectives:To assess the prevalence of traditional vaginal agent use in Malawian women and its association with HIV infection.Methods:Consenting, consecutive antenatal women were administered a questionnaire and screened for sexually transmitted diseases (STD) including HIV.Results:Of the 6603 consenting women, 886 (13%) reported using intravaginal agents for tightening and 2222 (34%) for self‐treatment of vaginal discharge and itching. A higher proportion of HIV‐infected than uninfected women (17% versus 14%) reported use of intravaginal agents for treatment (odds ratio, 1.29; 95% confidence interval, 1.05‐1.57), but no difference in HIV status was found when these agents were used for tightening. In multivariate analysis, vaginal agent use for treatment was independently associated with HIV seropositivity.Conclusions:The association of HIV infection with vaginal agents for self‐treatment, but not for tightening, suggests that STD may play a role or that vaginal agents are used differently for the two purposes. In addition to a small increased risk of HIV infection associated with vaginal agent use, these agents may interfere with condom effectiveness or acceptability of vaginal microbicides.