Screening autoantibody profiles in systemic rheumatic disease with a diagnostic protein microarray that uses a filtration-assisted nanodot array luminometric immunoassay (NALIA)

McBride, Jeffrey D. and Gabriel, Francis Guy and Fordham, John and Kolind, Torsten and Barcenas-Morales, Gabriela and Isenberg, David A. and Swana, Marlene and Delves, Peter J. and Lund, Torben and Cree, Ian A. and Roitt, Ivan (2008) Screening autoantibody profiles in systemic rheumatic disease with a diagnostic protein microarray that uses a filtration-assisted nanodot array luminometric immunoassay (NALIA). Clinical Chemistry, 54 (5). pp. 883-890. ISSN 0009-9147

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Abstract

Background: We developed a cost-efficient modular system for multiplex analysis of the multiple autoantibodies that characterize systemic rheumatoid diseases.

Methods: The nanodot array luminometric immunoassay (NALIA) system consists of conventional 96-well membrane-bottomed plates in which antigens or antibodies are adsorbed onto the underside of the membrane. Current arrays use a 5 × 5 format (25 dots/well), which allows 10 analytes to be measured in duplicate: double-stranded DNA (dsDNA), centromere protein B (CENP-B), PCNA, Sm, Sm ribonucleoprotein (Sm-RNP), U1-snRNP, Scl70, SSA/Ro, SSB/La, Jo-1, and controls. The test fluid, control sera, and subsequent reagents are drawn through the membrane. The captured analytes are quantified by monitoring chemiluminescence with a charge-coupled device (CCD) and analyzed with commercial array software.

Results: The assay can detect <20 × 103 IU/L of anti-dsDNA. The interwell CV was 10%–14%. There was an 83% concordance (κ = 0.56) between the NALIA results obtained for anti-dsDNA assayed by β-testing in a routine immunology diagnostic laboratory and the results obtained with a conventional ELISA reagent set. The concordance values for Ro, La, Sm, and RNP were 98% (κ, 0.92), 93% (κ, 0.41), 97% (κ, 0.62), and 97% (κ, 0.73), respectively.

Conclusion: The NALIA approach promises to provide a highly economical platform for a wide range of applications that require assays of multiple analytes. The degree of concordance of our results with a conventional reagent set was no less than that occurring between different commercial products. A sample of serum from a finger stick provides a volume sufficient to perform the array assay.

Item Type: Article
Research Areas: A. > School of Science and Technology > Natural Sciences
A. > School of Science and Technology > Natural Sciences > Biomarkers for Cancer group
Item ID: 11256
Useful Links:
Depositing User: Devika Mohan
Date Deposited: 11 Jul 2013 05:42
Last Modified: 13 Oct 2016 14:27
URI: http://eprints.mdx.ac.uk/id/eprint/11256

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