title: Broad spectrum reactivity versus subtype specificity-trade-offs in serodiagnosis of influenza A virus infections by competitive ELISA.
abstract: Avian influenza viruses (AIVs) of the H5 and H7 subtypes can cause substantial economic losses in the poultry industry and are a potential threat to public health. Serosurveillance of poultry populations is an important monitoring tool and can also be used for control of vaccination campaigns. The purpose of this study was to develop broadly reactive, yet subtype-specific competitive ELISAs (cELISAs) for the specific detection of antibodies to the notifiable AIV subtypes H5 and H7 as an alternative to the gold standard haemagglutination inhibition assay (HI). Broadly reacting monoclonal competitor antibodies (mAbs) and genetically engineered subtype H5 or H7 haemagglutinin antigen, expressed and in vivo biotinylated in insect cells, were used to develop the cELISAs. Sera from galliform species and water fowl (n=793) were used to evaluate the performance characteristics of the cELISAs. For the H5 specific cELISA, 98.1% test sensitivity and 91.5% test specificity (97.7% and 90.2% for galliforms; 98.9% and 92.6% for waterfowl), and for the H7 cELISA 97.3% sensitivity and 91.8% specificity (95.3% and 98.9% for galliforms; 100% and 82.7% for waterfowl) were reached when compared to HI. The use of competitor mAbs with broad spectrum reactivity within an AIV haemagglutinin subtype allowed for homogenous detection with high sensitivity of subtype-specific antibodies induced by antigenically widely distinct isolates including antigenic drift variants. However, a trade-off regarding sensitivity versus nonspecific detection of interfering antibodies induced by phylo- and antigenically closely related subtypes, e.g., H5 versus H2 and H7 versus H15, must be considered. The observed intersubtype antibody cross-reactivity remains a disturbance variable in AIV subtype-specific serodiagnosis which negatively affects specificity.
authors.affiliation: Institute of Diagnostic Virology, Friedrich-Loeffler-Institute, Federal Research Institute for Animal Health, Insel Riems, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Postel
authors.forename: M
authors.initials: M
authors.lastname: Ziller
authors.forename: M
authors.initials: M
authors.lastname: Rudolf
authors.forename: T
authors.initials: T
authors.lastname: Letzel
authors.forename: Ralf
authors.initials: R
authors.lastname: Ehricht
authors.forename: P
authors.initials: P
authors.lastname: Pourquier
authors.forename: M
authors.initials: M
authors.lastname: Dauber
authors.forename: C
authors.initials: C
authors.lastname: Grund
authors.forename: Martin
authors.initials: M
authors.lastname: Beer
authors.forename: T C
authors.initials: TC
authors.lastname: Harder
mesh_terms: D000818:Animals; D001717:Birds; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D009980:Influenza A virus / Q000276:immunology / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; C109963:hemagglutinin, avian influenza A virus