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A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays

Anti-nuclear antibodies (ANA) have traditionally been evaluated using indirect fluorescence assays (IFA) with HEp-2 cells. Quantitative immunoassays (EIA) have replaced the use of HEp-2 cells in some laboratories. Here, we evaluated ANA in 400 consecutive and unselected routinely referred patients u...

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Autores principales: Baronaite, Renata, Engelhart, Merete, Mørk Hansen, Troels, Thamsborg, Gorm, Slott Jensen, Hanne, Stender, Steen, Szecsi, Pal Bela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3926329/
https://www.ncbi.nlm.nih.gov/pubmed/24592328
http://dx.doi.org/10.1155/2014/534759
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author Baronaite, Renata
Engelhart, Merete
Mørk Hansen, Troels
Thamsborg, Gorm
Slott Jensen, Hanne
Stender, Steen
Szecsi, Pal Bela
author_facet Baronaite, Renata
Engelhart, Merete
Mørk Hansen, Troels
Thamsborg, Gorm
Slott Jensen, Hanne
Stender, Steen
Szecsi, Pal Bela
author_sort Baronaite, Renata
collection PubMed
description Anti-nuclear antibodies (ANA) have traditionally been evaluated using indirect fluorescence assays (IFA) with HEp-2 cells. Quantitative immunoassays (EIA) have replaced the use of HEp-2 cells in some laboratories. Here, we evaluated ANA in 400 consecutive and unselected routinely referred patients using IFA and automated EIA techniques. The IFA results generated by two independent laboratories were compared with the EIA results from antibodies against double-stranded DNA (dsDNA), from ANA screening, and from tests of the seven included subantigens. The final IFA and EIA results for 386 unique patients were compared. The majority of the results were the same between the two methods (n = 325, 84%); however, 8% (n = 30) yielded equivocal results (equivocal-negative and equivocal-positive) and 8% (n = 31) yielded divergent results (positive-negative). The results showed fairly good agreement, with Cohen's kappa value of 0.30 (95% confidence interval (CI) = 0.14–0.46), which decreased to 0.23 (95% CI = 0.06–0.40) when the results for dsDNA were omitted. The EIA method was less reliable for assessing nuclear and speckled reactivity patterns, whereas the IFA method presented difficulties detecting dsDNA and Ro activity. The automated EIA method was performed in a similar way to the conventional IFA method using HEp-2 cells; thus, automated EIA may be used as a screening test.
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spelling pubmed-39263292014-03-03 A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays Baronaite, Renata Engelhart, Merete Mørk Hansen, Troels Thamsborg, Gorm Slott Jensen, Hanne Stender, Steen Szecsi, Pal Bela Autoimmune Dis Research Article Anti-nuclear antibodies (ANA) have traditionally been evaluated using indirect fluorescence assays (IFA) with HEp-2 cells. Quantitative immunoassays (EIA) have replaced the use of HEp-2 cells in some laboratories. Here, we evaluated ANA in 400 consecutive and unselected routinely referred patients using IFA and automated EIA techniques. The IFA results generated by two independent laboratories were compared with the EIA results from antibodies against double-stranded DNA (dsDNA), from ANA screening, and from tests of the seven included subantigens. The final IFA and EIA results for 386 unique patients were compared. The majority of the results were the same between the two methods (n = 325, 84%); however, 8% (n = 30) yielded equivocal results (equivocal-negative and equivocal-positive) and 8% (n = 31) yielded divergent results (positive-negative). The results showed fairly good agreement, with Cohen's kappa value of 0.30 (95% confidence interval (CI) = 0.14–0.46), which decreased to 0.23 (95% CI = 0.06–0.40) when the results for dsDNA were omitted. The EIA method was less reliable for assessing nuclear and speckled reactivity patterns, whereas the IFA method presented difficulties detecting dsDNA and Ro activity. The automated EIA method was performed in a similar way to the conventional IFA method using HEp-2 cells; thus, automated EIA may be used as a screening test. Hindawi Publishing Corporation 2014 2014-01-28 /pmc/articles/PMC3926329/ /pubmed/24592328 http://dx.doi.org/10.1155/2014/534759 Text en Copyright © 2014 Renata Baronaite et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Baronaite, Renata
Engelhart, Merete
Mørk Hansen, Troels
Thamsborg, Gorm
Slott Jensen, Hanne
Stender, Steen
Szecsi, Pal Bela
A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays
title A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays
title_full A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays
title_fullStr A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays
title_full_unstemmed A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays
title_short A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays
title_sort comparison of anti-nuclear antibody quantification using automated enzyme immunoassays and immunofluorescence assays
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3926329/
https://www.ncbi.nlm.nih.gov/pubmed/24592328
http://dx.doi.org/10.1155/2014/534759
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