Understanding the at-risk population's perception of Chagas disease is essential for its effective prevention and control. A cross-sectional survey of knowledge, attitudes, and practices toward Chagas disease was conducted with Bolivians in a highly endemic region of Bolivia and Latin American migrants in London. In total, 175 participants completed the survey: 100 Bolivians in a highly endemic village in Santa Cruz, Bolivia and 75 Latin American migrants in London (of whom 31 were from Bolivia). All participants from the endemic village and all Bolivian migrants in London knew of Chagas disease, whereas only 25% of other Latin American migrants had heard of it (P <0.001). In London, Bolivians had more knowledge of Chagas disease than those from other Chagas-endemic countries. In Bolivia only, better understanding of Chagas disease was associated with educational attainment. Only 4% of participants overall were aware of the risk of vertical transmission. Few Latin American migrants in London had previously been tested for Chagas disease, and most were not aware of how to access testing. Migration and level of endemicity may shape individuals' understanding of as well as attitudes and practices toward Chagas disease. A better understanding of these factors can guide effective prevention and control program development in both endemic and non-endemic settings.