Introduction: Hepatitis E virus (HEV) is an emergent virus of global importance; it is the etiological agent of sporadic cases and outbreaks of hepatitis. The epidemiology of this infection in Colombiais unknown.Objective: To determine the seropositivity for hepatitis E virus in Colombia in cases with clinical diagnosis of viral hepatitis.Materials and methods: Serum samples from patients that were sent to the Instituto Nacional de Salud during the period 2005-2010 (group 1) and samples sent to the Laboratorio Departamental deSalud Pública de Antioquia during the 2008-2009 period were included in this study (group 2). Serum samples were analyzed by immunoassay with commercial kits.Results: From the 344 analyzed samples, 8.7% were positive for anti-HEV; the frequency of anti-HEV IgM was 1.74% (6/344) and the frequency of anti-HEV IgG was 7.5% (26/344). A difference in frequency of anti-HEV between group 1 (6.3%) and group 2 (1.3%) was observed. The cases were identified in nine departments of Colombia.Conclusions: This is the first study of hepatitis E virus infection in patients with diagnosis of hepatitis in Colombia. The frequency of anti-HEV described in this population of patients in Colombia is similar to that described in other Latin American countries like Brazil, Perú and Uruguay. Considering the results of this study, it could be necessary to include hepatitis E virus infection serological markers in the differential diagnosis of viral hepatitis in Colombia.