Hepatitis E virus is responsible for sporadic cases of acute, self-limited viral hepatitis not only in endemic but also in industrialized countries. In addition, some reports confirm that it can cause chronic infection and even cirrhosis in immunosuppressed and also in patients infected with HIV. There are few data about prevalence and incidence of HEV chronic infection in HIV-HEV coinfected individuals in Spain. The aim of this study is to investigate the prevalence of anti-HEV IgG in a representative sample of 448 patients infected with HIV and determine the role of age, gender, and CD4 counts in the detection of anti-HEV IgG antibodies in blood. In addition, the clinical features and ALT levels in relation to the presence of anti-HEV IgM and/or HEV-RNA in the blood of these patients were investigated. Anti-HEV IgG antibodies were detected in serum using a commercial enzyme immunoassay. All positive samples were studied further for the presence of anti-HEV IgM antibodies. In addition, HEV RNA was amplified by reverse transcriptase (RT)-nested PCR in all serum samples with IgM anti-HEV. The overall prevalence of anti-HEV IgG was 10.4% (45/448, 95% C.I. 7.2-12.8%). HEV-RNA was found in only one patient out of the 45 anti-HEV IgG positive samples studied. Regarding to gender and CD4 count, no difference in seroprevalence could be observed. This prevalence data suggest that patients infected with HIV can be considered a risk group for HEV infection and that chronic coinfection HEV-HIV seems to be a very rare event.