T. cruzi antigens and conventional human exosomal markers (CD9, CD63, CD81 and CD82) were found in plasma‐derived EVs from chronic Chagas disease patients. EVs from these patients contained both T. cruzi antigens and human exosomal markers. Future biomarker identification investigations in Chagas disease by establishing a feasible method for isolating total circulating EVs. Here, CD9 is linked to Chagas disease.