BACKGROUND Poliovirus (PV) antibody seroprevalence studies assess population immunity, verify an immunization program's performance and vaccine efficacy, and guide polio eradication strategy. Namibia experienced a polio outbreak among adults in 2006, yet population seroimmunity was unknown. METHODS We tested 2061 specimens from Namibian pregnant females aged 15-44 years for neutralizing antibody to PV types 1-3 (PV1-3); all females were sampled during the 2010 National HIV Sentinel Survey. We determined the proportion of females seropositive for PV antibody by 5-year age strata, and analyzed factors associated with seropositivity, including age, gravidity, human immunodeficiency virus (HIV) infection status, residence, and antiretroviral treatment, by log-binomial regression. RESULTS The seroprevalence was 94.6% for PV1, 97.0% for PV2, and 85.1% for PV3. HIV-positive females had significantly lower seroprevalence than HIV-negative females for PV1 (91.8% vs 95.3%; P<.01) and PV3 (80.0% vs 86.1%; P<.01) but not for PV2 (96.4% vs 97.1%; P=.3). The prevalence ratio of seropositivity for HIV-positive females versus HIV-negative females was 0.95 (95% confidence interval [CI], .92-.98) for PV1, 0.99 (95% CI, .97-1.01) for PV2, and 0.92 (95% CI, .87-.96) for PV3. CONCLUSIONS Despite relatively high PV seroprevalence, Namibia might remain at risk for a PV outbreak, particularly in lower-seroprevalence populations, such as HIV-positive females. Namibia should continue to maintain high routine polio vaccination coverage.