title: Indirect immunofluorescence assay for the simultaneous detection of antibodies against clinically important old and new world hantaviruses.
abstract: In order to detect serum antibodies against clinically important Old and New World hantaviruses simultaneously, multiparametric indirect immunofluorescence assays (IFAs) based on biochip mosaics were developed. Each of the mosaic substrates consisted of cells infected with one of the virus types Hantaan (HTNV), Puumala (PUUV), Seoul (SEOV), Saaremaa (SAAV), Dobrava (DOBV), Sin Nombre (SNV) or Andes (ANDV). For assay evaluation, serum IgG and IgM antibodies were analyzed using 184 laboratory-confirmed hantavirus-positive sera collected at six diagnostic centers from patients actively or previously infected with the following hantavirus serotypes: PUUV (Finland, n=97); SEOV (China, n=5); DOBV (Romania, n=7); SNV (Canada, n=23); ANDV (Argentina and Chile, n=52). The control panel comprised 89 sera from healthy blood donors. According to the reference tests, all 184 patient samples were seropositive for hantavirus-specific IgG (n=177; 96%) and/or IgM (n=131; 72%), while all control samples were tested negative. In the multiparametric IFA applied in this study, 183 (99%) of the patient sera were IgG and 131 (71%) IgM positive (accordance with the reference tests: IgG, 96%; IgM, 93%). Overall IFA sensitivity for combined IgG and IgM analysis amounted to 100% for all serotypes, except for SNV (96%). Of the 89 control sera, 2 (2%) showed IgG reactivity against the HTNV substrate, but not against any other hantavirus. Due to the high cross-reactivity of hantaviral nucleocapsid proteins, endpoint titrations were conducted, allowing serotype determination in >90% of PUUV- and ANDV-infected patients. Thus, multiparametric IFA enables highly sensitive and specific serological diagnosis of hantavirus infections and can be used to differentiate PUUV and ANDV infection from infections with Murinae-borne hantaviruses (e.g. DOBV and SEOV).
authors.affiliation: EUROIMMUN Medizinische Labordiagnostika AG, Luebeck, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Lederer
authors.forename: Erik
authors.initials: E
authors.lastname: Lattwein
authors.forename: Merle
authors.initials: M
authors.lastname: Hanke
authors.forename: Karen
authors.initials: K
authors.lastname: Sonnenberg
authors.forename: Winfried
authors.initials: W
authors.lastname: Stoecker
authors.forename: Åke
authors.initials: Å
authors.lastname: Lundkvist
authors.forename: Antti
authors.initials: A
authors.lastname: Vaheri
authors.forename: Olli
authors.initials: O
authors.lastname: Vapalahti
authors.forename: Paul K S
authors.initials: PK
authors.lastname: Chan
authors.forename: Heinz
authors.initials: H
authors.lastname: Feldmann
authors.forename: Daryl
authors.initials: D
authors.lastname: Dick
authors.forename: Jonas
authors.initials: J
authors.lastname: Schmidt-Chanasit
authors.forename: Paula
authors.initials: P
authors.lastname: Padula
authors.forename: Pablo A
authors.initials: PA
authors.lastname: Vial
authors.forename: Raluca
authors.initials: R
authors.lastname: Panculescu-Gatej
authors.forename: Cornelia
authors.initials: C
authors.lastname: Ceianu
authors.forename: Paul
authors.initials: P
authors.lastname: Heyman
authors.forename: Tatjana
authors.initials: T
authors.lastname: Avšič-Županc
authors.forename: Matthias
authors.initials: M
authors.lastname: Niedrig
mesh_terms: D000914:Antibodies, Viral / Q000032:analysis* / Q000276:immunology*; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D006476:Orthohantavirus / Q000276:immunology*; D018778:Hantavirus Infections / Q000276:immunology; D006801:Humans; D008856:Microscopy, Fluorescence
chemical_list: D000914:Antibodies, Viral