Abstract INTRODUCTION The prevalence, public health impact, and epidemiological characteristics of hepatitis E virus (HEV) are poorly understood in Brazil. METHODS Serum samples from 535 individuals from three rural Afro-descendant communities located in eastern Brazilian Amazon were collected in October 2015 and tested for presence of anti-HEV IgM and IgG antibodies. Serologically positive samples were also tested for HEV-RNA. RESULTS Two cases were confirmed for anti-HEV IgM (0.3 %) and two cases for anti-HEV IgG (0.3 %). No sample was positive for HEV-RNA. CONCLUSIONS Results indicated low prevalence of HEV infection in Afro-descendant rural communities from the eastern Brazilian Amazon.