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Biomolecules
Volume 16
Issue 5
10.3390/biom16050698
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Clinical Relevance and Methodological Comparison of Anti-MDA5 Antibody Detection: A Five-Year Retrospective and Exploratory Pilot Study

by
Sándor Mogyoróssy
1,2,
Gábor Nagy
3,
Zoltán Griger
4,
Melinda Nagy-Vincze
4,
Monika Bodoki
1,
Dóra Csige
1,
Tünde Tarr
4,
Éva Zöld
4,
György Pfliegler
5,
Boglárka Csilla Brúgós
5,
Hui Lu
6,
Sarah Tansley
6,
Péter Antal-Szalmás
3,
Andrea Domján
1,
Szilvia Szamosi
1,
Gabriella Szűcs
1,
Zoltán Szekanecz
1,
Ágnes Horváth
1,† and
Levente Bodoki
1,2,*,†
1
Department of Rheumatology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary
2
Doctoral School of Medical Sciences, University of Debrecen, 4032 Debrecen, Hungary
3
Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary
4
Division of Clinical Immunology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary
5
Centre of Rare Diseases, Department of Internal Medicine, University of Debrecen, 4032 Debrecen, Hungary
6
Department of Life Sciences, University of Bath, Bath BA2 7AY, UK
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Biomolecules 2026, 16(5), 698; https://doi.org/10.3390/biom16050698
Submission received: 20 March 2026 / Revised: 24 April 2026 / Accepted: 7 May 2026 / Published: 8 May 2026
(This article belongs to the Section Molecular Medicine)
Versions Notes

Abstract

Anti-melanoma differentiation-associated gene 5 (anti-MDA5) antibodies are critical biomarkers in myositis, associated with distinct clinical features and prognosis. This study aimed to evaluate the proportion of anti-MDA5 positivity and compare the diagnostic performance of local immunoblotting (IB) with gold-standard immunoprecipitation (IP). We performed a retrospective analysis of 3272 physician-requested anti-MDA5 IB determinations over a five-year period (2019–2023). A subsequent exploratory pilot study of ten Hungarian patients with myositis was conducted to compare IB results with radiolabeled protein IP. Confirmatory in-house enzyme-linked immunosorbent assay (ELISA) was used to distinguish between 140 kDa bands (anti-MDA5 vs. anti-NXP2). Indirect immunofluorescence (IIF) on HEp-2 cells was also evaluated. In the retrospective cohort, 3.7% (n = 121) of samples were non-negative. Among 64 borderline patients, only one (1.6%) had a definitive diagnosis of dermatomyositis (DM). Conversely, the proportion of confirmed myositis cases was notably higher among patients with strong positive IB results. In our exploratory cross-sectional pilot study, complete concordance between the two assays was observed for negative and strong positive results. Discrepancies were noted in borderline and weak positivity ranges, where anti-MDA5 was not detected by IP; instead, alternative autoantibodies were identified. The three IP-confirmed MDA5 positive samples were all validated by ELISA. The characteristic IIF cytoplasmic staining was identifiable in 2 out of 10 cases (20%). In our cohort, borderline IB cases were frequently potential false positives, highlighting the need for careful clinical evaluation. Borderline and weak results require clinical correlation or confirmatory testing to avoid misdiagnosis.
Keywords: anti-MDA5; dermatomyositis; idiopathic inflammatory myopathy; immunoblot; immunoprecipitation anti-MDA5; dermatomyositis; idiopathic inflammatory myopathy; immunoblot; immunoprecipitation

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MDPI and ACS Style

Mogyoróssy, S.; Nagy, G.; Griger, Z.; Nagy-Vincze, M.; Bodoki, M.; Csige, D.; Tarr, T.; Zöld, É.; Pfliegler, G.; Brúgós, B.C.; et al. Clinical Relevance and Methodological Comparison of Anti-MDA5 Antibody Detection: A Five-Year Retrospective and Exploratory Pilot Study. Biomolecules 2026, 16, 698. https://doi.org/10.3390/biom16050698

AMA Style

Mogyoróssy S, Nagy G, Griger Z, Nagy-Vincze M, Bodoki M, Csige D, Tarr T, Zöld É, Pfliegler G, Brúgós BC, et al. Clinical Relevance and Methodological Comparison of Anti-MDA5 Antibody Detection: A Five-Year Retrospective and Exploratory Pilot Study. Biomolecules. 2026; 16(5):698. https://doi.org/10.3390/biom16050698

Chicago/Turabian Style

Mogyoróssy, Sándor, Gábor Nagy, Zoltán Griger, Melinda Nagy-Vincze, Monika Bodoki, Dóra Csige, Tünde Tarr, Éva Zöld, György Pfliegler, Boglárka Csilla Brúgós, and et al. 2026. "Clinical Relevance and Methodological Comparison of Anti-MDA5 Antibody Detection: A Five-Year Retrospective and Exploratory Pilot Study" Biomolecules 16, no. 5: 698. https://doi.org/10.3390/biom16050698

APA Style

Mogyoróssy, S., Nagy, G., Griger, Z., Nagy-Vincze, M., Bodoki, M., Csige, D., Tarr, T., Zöld, É., Pfliegler, G., Brúgós, B. C., Lu, H., Tansley, S., Antal-Szalmás, P., Domján, A., Szamosi, S., Szűcs, G., Szekanecz, Z., Horváth, Á., & Bodoki, L. (2026). Clinical Relevance and Methodological Comparison of Anti-MDA5 Antibody Detection: A Five-Year Retrospective and Exploratory Pilot Study. Biomolecules, 16(5), 698. https://doi.org/10.3390/biom16050698

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