Saarinen, Niila V. V.Lehtonen, JussiVeijola, RiittaLempainen, JohannaKnip, MikaelHyöty, HeikkiLaitinen, Olli H.Hytönen, Vesa P.2022-09-282022-09-282020-06-26Saarinen, N.V.V.; Lehtonen, J.; Veijola, R.; Lempainen, J.; Knip, M.; Hyöty, H.; Laitinen, O.H.; Hytönen, V.P. Multiplexed High-Throughput Serological Assay for Human Enteroviruses. Microorganisms 2020, 8, 963.http://hdl.handle.net/10138/348753Immunological assays detecting antibodies against enteroviruses typically use a single enterovirus serotype as antigen. This limits the ability of such assays to detect antibodies against different enterovirus types and to detect possible type-specific variation in antibody responses. We set out to develop a multiplexed assay for simultaneous detection of antibodies against multiple enterovirus and rhinovirus types encompassing all human infecting species. Seven recombinant VP1 proteins from enteroviruses EV-A to EV-D and rhinoviruses RV-A to RV-C species were produced. Using Meso Scale Diagnostics U-PLEX platform we were able to study antibody reactions against these proteins as well as non-structural enterovirus proteins in a single well with 140 human serum samples. Adults had on average 33-fold stronger antibody responses to these antigens (p < 10−11) compared to children, but children had less cross-reactivity between different enterovirus types. The results suggest that this new high-throughput assay offers clear benefits in the evaluation of humoral enterovirus immunity in children, giving more exact information than assays that are based on a single enterovirus type as antigen.eng2022-09-28http://purl.org/eprint/entityType/JournalArticle