title: Specific detection of antibodies to different flaviviruses using a new immune complex ELISA.
abstract: Immune complex (IC) ELISAs for IgG antibodies to various virus antigens have turned out to be both highly specific and sensitive. During incubation of a labelled antigen with the serum samples, ICs are formed, which bind to microtiter plates coated with rheumatoid factor (RF) IgM. Here, we describe an improved coating of the solid-phase support comparing various Fc-receptor molecules. IC ELISAs were applied to detect human IgG antibodies to the highly virus-specific ED3 domain of West Nile- and tick-borne encephalitis virus envelopes. Compared with other Fc-receptor molecules like RF or C1q, FcγRIIA (CD32) turned out to bind the ICs composed of IgG antibodies and peroxidase-labelled ED3 antigens more efficiently. Due to low background reactions, sera could be tested at a dilution of 1:10. Moreover, using CD32 instead of RF coating, anti-flavivirus antibodies could be detected in various animal species.
authors.affiliation: Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Str.74, 20359 Hamburg, Germany. Schmitz@bni-hamburg.de
authors.forename: Herbert
authors.initials: H
authors.lastname: Schmitz
authors.forename: Martin
authors.initials: M
authors.lastname: Gabriel
authors.forename: Petra
authors.initials: P
authors.lastname: Emmerich
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D004669:Encephalitis Viruses, Tick-Borne / Q000276:immunology / Q000472:pathogenicity; D004675:Encephalitis, Tick-Borne / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D017452:Receptors, IgG / Q000276:immunology; D012217:Rheumatoid Factor / Q000276:immunology; D012680:Sensitivity and Specificity; D014611:Vaccination; D014759:Viral Envelope Proteins / Q000276:immunology; D014901:West Nile Fever / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D014902:West Nile virus / Q000276:immunology / Q000472:pathogenicity
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D007074:Immunoglobulin G; D017452:Receptors, IgG; D014759:Viral Envelope Proteins; C065696:glycoprotein E, Flavivirus; D012217:Rheumatoid Factor