Strongyloides stercoralis, a pathogenic roundworm, is considered endemic in several tropical and subtropical areas worldwide. Indigenous populations have the highest soil-transmitted helminthiases-related mortality rates, but the prevalence and risk factors associated withS.stercoralisin Brazilian indigenous populations have not been established. Thus, the present study aimed to assess the seroprevalence and associated risk factors forS.stercoralisin indigenous communities and the healthcare professionals serving them in Brazil. Indigenous populations living in nine communities and healthcare professionals were tested for anti-S.stercoralisantibodies by ELISA. A questionnaire was used to assess socio-epidemiological information. Associated risk factors for seropositivity were tested by chi-square or Fisher’s exact tests, using univariate analyses and multivariate logistic regression. Overall, 174/463 (37.6%; CI 95%: 33.3–42.1) indigenous persons and 77/147 (52.4%; 95% CI: 44.3–60.3) healthcare professionals were seropositive for anti-S.stercoralisantibodies. Seropositivity among the two groups was statistically significant (p = 0.0016; OR = 0.547; 95% CI: 0.376–0.796) and revealed that healthcare professionals were 1.83 times more likely to be seropositive. The multivariate analysis showed that being male or being adult were also risk factors, while having a septic tank as a sanitary facility represented a protective factor forS.stercoralisexposure in indigenous persons. None of the variables evaluated were associated withS.stercoralisexposure in the professional group. The study herein has reported a high seroprevalence toStrongyloides stercoralisin indigenous communities of Brazil and healthcare professionals, warning for potential public health concerns of strongyloidiasis in such populations.