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

Article

McBride, J., Gabriel, F., Fordham, J., Kolind, T., Barcenas-Morales, G., Isenberg, D., Swana, M., Delves, P., Lund, T., Cree, I. and Roitt, I. 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. https://doi.org/10.1373/clinchem.2007.098418
TypeArticle
TitleScreening autoantibody profiles in systemic rheumatic disease with a diagnostic protein microarray that uses a filtration-assisted nanodot array luminometric immunoassay (NALIA)
AuthorsMcBride, J., Gabriel, F., Fordham, J., Kolind, T., Barcenas-Morales, G., Isenberg, D., Swana, M., Delves, P., Lund, T., Cree, I. and Roitt, I.
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.

Research GroupBiomarkers for Cancer group
PublisherAmerican Association for Clinical Chemistry
JournalClinical Chemistry
ISSN0009-9147
Publication dates
Print01 May 2008
Publication process dates
Deposited11 Jul 2013
Output statusPublished
Digital Object Identifier (DOI)https://doi.org/10.1373/clinchem.2007.098418
LanguageEnglish
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