title: A novel indirect immunofluorescence test for the detection of IgG and IgA antibodies for diagnosis of Hepatitis E Virus infections.
abstract: Hepatitis E Virus (HEV) causes epidemic infections in regions of poor hygiene in the developing world. Over the last years, however, increasing numbers of autochthonous infections in industrialized countries have been described, leading to new interest in this pathogen. Currently available serological test formats to detect IgG and IgM antibodies are mainly based on bacterially expressed ORF2 and ORF3 antigens and often give ambiguous results. The objective of this study was the development of a different assay format for HEV diagnosis--a HEV immunofluorescence test (HEV-IFT) based on mammalian cells transiently expressing recombinant HEV ORF2 protein with a simple production and staining protocol and the investigation of its performance and methodical feasibility under diagnostic laboratory conditions. 31 sera of patients at different phases of HEV infection and 40 control sera from a non-endemic region were analyzed for anti-HEV IgG, IgM, and IgA antibodies. The HEV-IFT detected successfully anti-HEV IgG and IgA, but not anti-HEV IgM antibodies. In the study group the HEV-IFT was able to confirm HEV infections and to support diagnosis when ambiguous results were obtained by commercial assays. Signal localization and staining patterns helped to gather additional information about reactive antibodies present in patient sera. In conclusion the developed IFT for the detection of anti-HEV IgG and IgA antibodies can be used for diagnosis and for the serological confirmation of HEV infections.
authors.affiliation: Max von Pettenkofer-Institute, Virology, Ludwig-Maximilians-University of Munich, Pettenkoferstrasse 9a, D-80336 Munich, Germany. osterman@mvp.uni-muenchen.de
authors.forename: Andreas
authors.initials: A
authors.lastname: Osterman
authors.forename: Maria G
authors.initials: MG
authors.lastname: Vizoso-Pinto
authors.forename: Jette
authors.initials: J
authors.lastname: Jung
authors.forename: Gundula
authors.initials: G
authors.lastname: Jaeger
authors.forename: Josef
authors.initials: J
authors.lastname: Eberle
authors.forename: Hans
authors.initials: H
authors.lastname: Nitschko
authors.forename: Armin
authors.initials: A
authors.lastname: Baiker
mesh_terms: D019411:Clinical Laboratory Techniques / Q000379:methods*; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D006508:Hepatitis Antibodies / Q000097:blood*; D016751:Hepatitis E / Q000175:diagnosis*; D016752:Hepatitis E virus / Q000276:immunology*; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood*; D007074:Immunoglobulin G / Q000097:blood*
chemical_list: D006508:Hepatitis Antibodies; D007070:Immunoglobulin A; D007074:Immunoglobulin G