title: Array-in-well serodiagnostic assay utilizing upconverting phosphor label technology.
abstract: In this study, a multiplex serological array-in-well assay was constructed for simultaneous detection of serum IgG antibodies against parvovirus B19 and human adenovirus. The array was prepared in streptavidin-coated 96-well microtiter plates by spotting biotinylated parvovirus B19 virus-like-particles, adenovirus type 2 and 5 hexon antigens, negative control of human serum albumin and positive controls of human IgG and anti-human IgG antibodies on the bottom of each well in an array format with a printable area of 2 mm × 2 mm. The array-in-well assay was evaluated with serum samples (n=89) of different antibody status as determined by commercial enzyme immunoassay for parvovirus IgG, and by in-house enzyme immunoassay for adenovirus IgG. The bound serum anti-parvovirus IgG, anti-adenovirus IgG, and total IgG antibodies were detected with anti-human IgG antibody coated photon upconverting nanoparticles and the assay was measured with an anti-Stokes photoluminescence imager. Detection of specific antibodies by the multiplex array-in-well assay was in good agreement (100% for parvovirus B19 and 96% for adenovirus) with the reference results. In conclusion, the array-in-well with upconverting phosphor reporter technology was able to detect antiviral antibodies in human sera, and represents an efficient serodiagnostic concept that is a promising new tool for multiplex serology.
authors.affiliation: Department of Virology, University of Turku, Kiinamyllynkatu 13, FI-20520 Turku, Finland; Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland. Electronic address: minna.yliharsila@utu.fi.
authors.forename: Minna
authors.initials: M
authors.lastname: Ylihärsilä
authors.affiliation: Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Siina
authors.initials: S
authors.lastname: Alaranta
authors.affiliation: Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Susanne
authors.initials: S
authors.lastname: Lahdenperä
authors.affiliation: Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Satu
authors.initials: S
authors.lastname: Lahtinen
authors.affiliation: Haartman Institute, Haartmaninkatu 3, University of Helsinki and HUSLAB, FI-00290 Helsinki, Finland.
authors.forename: Benedict
authors.initials: B
authors.lastname: Arku
authors.affiliation: Haartman Institute, Haartmaninkatu 3, University of Helsinki and HUSLAB, FI-00290 Helsinki, Finland.
authors.forename: Klaus
authors.initials: K
authors.lastname: Hedman
authors.affiliation: Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland.
authors.forename: Tero
authors.initials: T
authors.lastname: Soukka
authors.affiliation: Department of Virology, University of Turku, Kiinamyllynkatu 13, FI-20520 Turku, Finland.
authors.forename: Matti
authors.initials: M
authors.lastname: Waris
mesh_terms: D000258:Adenovirus Infections, Human / Q000175:diagnosis*; D000260:Adenoviruses, Human / Q000276:immunology*; D000328:Adult; D000914:Antibodies, Viral / Q000097:blood*; D002675:Child, Preschool; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D008163:Luminescent Measurements; D010322:Parvoviridae Infections / Q000175:diagnosis*; D016732:Parvovirus B19, Human / Q000276:immunology*; D012698:Serologic Tests / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D007074:Immunoglobulin G
keywords: Adenovirus; Array-in-well; Multiplex; Parvovirus B19; Serodiagnostic; Upconverting phosphor