ArticleFrontiers in immunology2026
A non-radioactive modified immunoprecipitation assay for diagnostic detection of anti-MDA5 antibodies in idiopathic inflammatory myopathies.
Article in Frontiers in immunology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Introduction: Anti-melanoma differentiation-associated gene 5 (MDA5) autoantibodies are clinically important in idiopathic inflammatory myopathies (IIM), but currently used screening assays may yield clinically misleading false-positive results. We aimed to compare commonly used anti-MDA5 detection methods and to evaluate a modified, non-radioactive immunoprecipitation assay as a practical confirmatory method for anti-MDA5 testing. Methods: We analysed 93 individuals, including 55 patients with IIM, 18 non-IIM cases with line-blot anti-MDA5 positivity, and 20 healthy controls. Anti-MDA5 antibodies were assessed using line blot assay, immunocytochemistry, radioimmunoassay and a modified immunoprecipitation assay using activated THP-1 cell lysates. Assay performance and agreement were evaluated using radioimmunoassay as the reference method. Results: Radioimmunoassay identified substantial discrepancies in anti-MDA5 classification by routine screening assays. Among 55 patients with IIM, 5 of 37 cases initially classified as MDA5-positive by line blot (13.5%) were false positives, and all 18 non-IIM line-blot anti-MDA5-positive cases were likewise unconfirmed by radioimmunoassay. Across all 93 individuals, line blot and immunocytochemistry showed false-positive rates of 38.33% and 21.67%, respectively. In contrast, the modified immunoprecipitation assay showed complete concordance with radioimmunoassay (kappa = 1.0), with no false-positive or false-negative results. Conclusion: False-positive anti-MDA5 results are common with line blot and immunocytochemistry, particularly in clinically ambiguous cases. The modified immunoprecipitation assay showed complete concordance with radioimmunoassay in this cohort and may provide a clinically useful non-radioactive confirmatory approach for anti-MDA5 antibody detection.
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