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The reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity
OBJECTIVES: In order to address the reliability of commercial assays to identify myositis-specific and -associated autoantibodies, we aimed to compare the results of two commercial immunoassays with the results obtained by protein immunoprecipitation. METHODS: Autoantibody status was determined usin...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7382594/ https://www.ncbi.nlm.nih.gov/pubmed/32030410 http://dx.doi.org/10.1093/rheumatology/keaa021 |
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author | Tansley, Sarah L Li, Danyang Betteridge, Zoe E McHugh, Neil J |
author_facet | Tansley, Sarah L Li, Danyang Betteridge, Zoe E McHugh, Neil J |
author_sort | Tansley, Sarah L |
collection | PubMed |
description | OBJECTIVES: In order to address the reliability of commercial assays to identify myositis-specific and -associated autoantibodies, we aimed to compare the results of two commercial immunoassays with the results obtained by protein immunoprecipitation. METHODS: Autoantibody status was determined using radio-labelled protein immunoprecipitation for patients referred to our laboratory for myositis autoantibody characterization. For each autoantibody of interest, the sera from 25 different patients were analysed by line blot (Euroline Myositis Antigen Profile 4, EuroImmun, Lübeck, Germany) and dot blot (D-Tek BlueDiver, Diagnostic Technology, Belrose, NSW, Australia). Sera from 134 adult healthy controls were analysed. RESULTS: Overall commercial assays performed reasonably well, with high agreement (Cohen’s κ >0.8). Notable exceptions were the detection of rarer anti-synthetases with κ < 0.2 and detection of anti-TIF1γ, where κ was 0.70 for the line blot and 0.31 for dot blot. Further analysis suggested that the proportion of patients with anti-TIF1γ may recognize a conformational epitope, limiting the ability of blotting-based assays that utilize denatured antigen to detect this clinically important autoantibody. A false-positive result occurred in 13.7% of samples analysed by line blot and 12.1% analysed by dot blot. CONCLUSION: The assays analysed do not perform well for all myositis-specific and -associated autoantibodies and overall false positives are relatively common. It is crucial that clinicians are aware of the limitations of the methods used by their local laboratory. Results must be interpreted within the clinical context and immunoprecipitation should still be considered in selected cases, such as apparently autoantibody-negative patients where anti-synthetase syndrome is suspected. |
format | Online Article Text |
id | pubmed-7382594 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-73825942020-07-29 The reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity Tansley, Sarah L Li, Danyang Betteridge, Zoe E McHugh, Neil J Rheumatology (Oxford) Clinical Science OBJECTIVES: In order to address the reliability of commercial assays to identify myositis-specific and -associated autoantibodies, we aimed to compare the results of two commercial immunoassays with the results obtained by protein immunoprecipitation. METHODS: Autoantibody status was determined using radio-labelled protein immunoprecipitation for patients referred to our laboratory for myositis autoantibody characterization. For each autoantibody of interest, the sera from 25 different patients were analysed by line blot (Euroline Myositis Antigen Profile 4, EuroImmun, Lübeck, Germany) and dot blot (D-Tek BlueDiver, Diagnostic Technology, Belrose, NSW, Australia). Sera from 134 adult healthy controls were analysed. RESULTS: Overall commercial assays performed reasonably well, with high agreement (Cohen’s κ >0.8). Notable exceptions were the detection of rarer anti-synthetases with κ < 0.2 and detection of anti-TIF1γ, where κ was 0.70 for the line blot and 0.31 for dot blot. Further analysis suggested that the proportion of patients with anti-TIF1γ may recognize a conformational epitope, limiting the ability of blotting-based assays that utilize denatured antigen to detect this clinically important autoantibody. A false-positive result occurred in 13.7% of samples analysed by line blot and 12.1% analysed by dot blot. CONCLUSION: The assays analysed do not perform well for all myositis-specific and -associated autoantibodies and overall false positives are relatively common. It is crucial that clinicians are aware of the limitations of the methods used by their local laboratory. Results must be interpreted within the clinical context and immunoprecipitation should still be considered in selected cases, such as apparently autoantibody-negative patients where anti-synthetase syndrome is suspected. Oxford University Press 2020-08 2020-02-06 /pmc/articles/PMC7382594/ /pubmed/32030410 http://dx.doi.org/10.1093/rheumatology/keaa021 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of the British Society for Rheumatology. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Clinical Science Tansley, Sarah L Li, Danyang Betteridge, Zoe E McHugh, Neil J The reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity |
title | The reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity |
title_full | The reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity |
title_fullStr | The reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity |
title_full_unstemmed | The reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity |
title_short | The reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity |
title_sort | reliability of immunoassays to detect autoantibodies in patients with myositis is dependent on autoantibody specificity |
topic | Clinical Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7382594/ https://www.ncbi.nlm.nih.gov/pubmed/32030410 http://dx.doi.org/10.1093/rheumatology/keaa021 |
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