Hepatitis-E-virus-associated antigen: improved detection in stools by protein Fv removal

ELISA detection of a hepatitis-E-virus-associated antigen (HEV-AAg) in stools was reappraised for its possible interference with a new Fab-binding factor, termed protein Fv, released during infectious hepatitis. Transaminase elevation, HEV-AAg discharge and Fv leakage appeared simultaneously in a Cercopithecus monkey inoculated with infected stools. Labelled normal, or immune human IgG, were compared with pre- and post-inoculation simian IgG, for HEV-AAg and Fv detection. Coated normal and patient human IgM were also compared to pre- and post-inoculation simian IgM in HEV-AAg and Fv capture assays. Simian IgM and beta-galactosidase-labelled simian IgG minimized Fv interference and appeared to be the best adapted system for HEV-AAg detection. Nevertheless, Fv was still the cause of false-positive interpretations in some cases; therefore adsorption with monoclonal IgM was required to ensure HEV specificity. The improved test was performed on stools from 30 Senegalese patients hospitalized for various sporadic attacks of viral hepatitis. HEV-AAg was detected in 6 out of 30 cases and no positivity was observed in patients suffering from hepatitis due to HAV, HBV, cytomegalovirus or Epstein-Barr virus. The specificity of the assay was confirmed by inhibition experiments with the sera from HEV-infected patients. Hence, this inhibition assay can also be used to detect serum antibodies to HEV-AAg.