title: Immunoassay for serologic diagnosis of influenza type A using recombinant DNA produced nucleoprotein antigen and monoclonal antibody to human IgG.
abstract: Influenza type A nucleoprotein (NP) derived from the full length cloned gene expressed in E. coli was evaluated in a solid phase enzyme immunoassay (EIA) for detection of human antibody to influenza. Monoclonal antibody to human IgG was used for detection. Direct and indirect assays were developed and sera were tested in serial and single dilution formats. Preliminary results indicated that recombinant-and virion-derived NP antigens were comparable in binding ability to plastic and binding human antibody. Eighty-seven paired sera from influenza patients were tested. The most sensitive assay (indirect-serial dilution) detected 56 (64%) rises and the simplest assay (direct-single dilution) detected 43 (49%) rises, compared to 36 (41%) for complement fixation. Paired sera from 18 control patients showed no evidence of antibody rises by any of the assays. Forty-nine paired sera from influenza B infected patients were negative for antibody rises except for one borderline rise by the indirect-serial dilution assay. These results indicate that the use of recombinant DNA derived nucleoprotein for immunoassay is feasible. The sensitivity of immunoassays using NP adsorbed to the solid phase and monoclonal antibody specific for human IgG to detect bound antibody exceeded that of conventional complement fixation testing for establishing serologic evidence of influenza type A infection.
authors.affiliation: Influenza Branch, Centers for Disease Control, Atlanta, Georgia 30333.
authors.forename: M W
authors.initials: MW
authors.lastname: Harmon
authors.forename: I
authors.initials: I
authors.lastname: Jones
authors.forename: M
authors.initials: M
authors.lastname: Shaw
authors.forename: W
authors.initials: W
authors.lastname: Keitel
authors.forename: C B
authors.initials: CB
authors.lastname: Reimer
authors.forename: P
authors.initials: P
authors.lastname: Halonen
authors.forename: A P
authors.initials: AP
authors.lastname: Kendal
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D015153:Blotting, Western; D003168:Complement Fixation Tests; D004591:Electrophoresis, Polyacrylamide Gel; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D007124:Immunoenzyme Techniques; D009980:Influenza A virus / Q000276:immunology* / Q000302:isolation & purification; D007251:Influenza, Human / Q000175:diagnosis* / Q000276:immunology; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D014758:Viral Core Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D011994:Recombinant Proteins; D014758:Viral Core Proteins