Shame may increase HIV risk among stigmatized populations. The Personal Feelings Questionnaire-2 (PFQ-2) measures shame, but has not been validated in Spanish-speaking or nonclinical stigmatized populations disproportionately affected by HIV in resource-limited settings. We examined the psychometric properties of the Spanish-translated PFQ-2 shame subscale among female sex workers in two Mexico-U.S. border cities. From 2016 to 2017, 602 HIV-negative female sex workers in Tijuana and Ciudad Juarez participated in an efficacy trial evaluating a behavior change maintenance intervention. Interviewer-administered surveys collected information on shame (10-item PFQ-2 subscale), psychosocial factors, and sociodemographics. Item performance, confirmatory factor analysis, internal consistency, differential item functioning by city, and concurrent validity were assessed. Response options were collapsed to 3-point responses to improve item performance, and one misfit item was removed. The revised 9-item shame subscale supported a single construct and had good internal consistency (Cronbach's α = .86). Notable differential item functioning was found but resulted in a negligible effect on overall scores. Correlations between the revised shame subscale and guilt (r = .79, p < .01), depression (r = .69, p < .01), and emotional support (r = -.28, p < .01) supported concurrent validity. The revised PFQ-2 shame subscale showed good reliability and concurrent validity in our sample, and should be explored in other stigmatized populations.