Surface-activated microtiter-plate microarray for simultaneous CRP quantification and viral antibody detection |
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Authors: | Sari M Viitala Anne J Jääskeläinen Eira Kelo Helena Sirola Kirsi Moilanen Jukka Suni Antti Vaheri Olli Vapalahti Ale Närvänen |
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Institution: | 1. School of Pharmacy, University of Eastern Finland, Kuopio, Finland;2. Biocenter Kuopio, Kuopio, Finland;3. Haartman Institute, Department of Virology, University of Helsinki, Finland;4. HUSLAB, Department of Virology and Immunology, Helsinki University Central Hospital, Helsinki, Finland;5. Reagena Ltd, Finland;6. Mobidiag Ltd, Finland;g Faculty of Veterinary Medicine, Department of Basic Veterinary Biosciences, University of Helsinki, Helsinki, Finland |
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Abstract: | Microarrays are widely used in high-throughput DNA and RNA hybridization tests and recently adopted to protein and small molecule interaction studies in basic research and diagnostics. Parallel detection of serum antibodies and antigens has several potential applications in epidemiologic research, vaccine development, and in the diagnosis of allergies, autoimmunity, and infectious diseases. This study demonstrates an immobilization method for immunoassay-based microarray in conventional 96-well polystyrene plates for a serologic diagnostic method combined with quantitative C-reactive protein (CRP) assay. A synthetic peptide (HIV-1), a recombinant protein (Puumala hantavirus nucleocapsid), and purified virus preparations (Sindbis and adenoviruses) were used as antigens for virus-specific antibody detection and monoclonal anti-CRP antibody for antigen detection. The microarray was based on conventional enzyme immunoassays and densitometry from photographed results. Peptide and recombinant antigens functioned well, while whole virus antigens gave discrepant results in 1 out of 23 samples from the reference method, tested with human sera with various antibody responses. The CRP results were in concordance in the concentration range 0.5–150 mg/L with 2 commercially available CRP assays: ReaScan rapid test (R2 = 0.9975) and Cobas 6000 analyzer (R2 =0.9595). The results indicate that microtiter plates provide a promising platform for further development of microarrays for parallel antibody and antigen detection. |
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Keywords: | Microarray Serology Antigen detection |
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