Array-in-well serodiagnostic assay utilizing upconverting phosphor label technology.

Autor: Ylihärsilä M; 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., Alaranta S; Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland., Lahdenperä S; Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland., Lahtinen S; Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland., Arku B; Haartman Institute, Haartmaninkatu 3, University of Helsinki and HUSLAB, FI-00290 Helsinki, Finland., Hedman K; Haartman Institute, Haartmaninkatu 3, University of Helsinki and HUSLAB, FI-00290 Helsinki, Finland., Soukka T; Department of Biotechnology, University of Turku, Tykistökatu 6A, FI-20520 Turku, Finland., Waris M; Department of Virology, University of Turku, Kiinamyllynkatu 13, FI-20520 Turku, Finland.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2015 Sep 15; Vol. 222, pp. 224-30. Date of Electronic Publication: 2015 May 28.
DOI: 10.1016/j.jviromet.2015.05.012
Abstrakt: 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.
(Copyright © 2015. Published by Elsevier B.V.)
Databáze: MEDLINE
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