title: Competitive Homogeneous Immunoassay for Rapid Serodiagnosis of Hantavirus Disease.
abstract: In this study, we describe a competitive homogeneous immunoassay that makes use of Förster resonance energy transfer (FRET) in rapid detection of pathogen-specific antibodies. The assay principle is based on competition between a monoclonal antibody (MAb) and serum antibodies to a given antigen. In the assay, named competitive FRET immunoassay (CFRET-IA), the FRET signal is induced if MAb carrying a donor label binds to an acceptor-labeled antigen. Specific antibodies in serum compete for antigen binding, resulting in reduced FRET signal. The proof-of-principle for the assay was obtained using donor-labeled Puumala virus nucleocapsid protein (PUUV-N) and acceptor-labeled anti-PUUV-N MAb. The assay was evaluated by analyzing 329 clinical samples comprising 101 from individuals with acute PUUV infection, 42 from individuals with past infection, and 186 from individuals with PUUV-seronegative sera, and the results were compared to those of reference tests. The rapid serodiagnostic test we introduced herein performed with 100% sensitivity and 99% specificity for diagnosing acute hantavirus disease.
authors.affiliation: Medicum, Virology, University of Helsinki, Helsinki, Finland satu.hepojoki@helsinki.fi.
authors.forename: Satu
authors.initials: S
authors.lastname: Hepojoki
authors.affiliation: Medicum, Virology, University of Helsinki, Helsinki, Finland.
authors.forename: Juuso
authors.initials: J
authors.lastname: Rusanen
authors.affiliation: Medicum, Virology, University of Helsinki, Helsinki, Finland.
authors.forename: Jussi
authors.initials: J
authors.lastname: Hepojoki
authors.affiliation: Medicum, Virology, University of Helsinki, Helsinki, Finland.
authors.forename: Visa
authors.initials: V
authors.lastname: Nurmi
authors.affiliation: Medicum, Virology, University of Helsinki, Helsinki, Finland Helsinki University Hospital, Helsinki, Finland.
authors.forename: Antti
authors.initials: A
authors.lastname: Vaheri
authors.affiliation: University of Uppsala, Uppsala, Sweden.
authors.forename: Åke
authors.initials: Å
authors.lastname: Lundkvist
authors.affiliation: Medicum, Virology, University of Helsinki, Helsinki, Finland Helsinki University Hospital, Helsinki, Finland.
authors.forename: Klaus
authors.initials: K
authors.lastname: Hedman
authors.affiliation: Medicum, Virology, University of Helsinki, Helsinki, Finland Helsinki University Hospital, Helsinki, Finland Veterinary Biosciences, University of Helsinki, Helsinki, Finland.
authors.forename: Olli
authors.initials: O
authors.lastname: Vapalahti
mesh_terms: D000914:Antibodies, Viral / Q000097:blood*; D031541:Fluorescence Resonance Energy Transfer; D006476:Orthohantavirus / Q000276:immunology*; D018778:Hantavirus Infections / Q000175:diagnosis*; D006801:Humans; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D013997:Time Factors
chemical_list: D000914:Antibodies, Viral