ArticleEuropean journal of medical research2025
Development of a novel flow cytometry multiplex immunoassay for the diagnosis of celiac disease simultaneously with autoimmune diabetes.
Article in European journal of medical research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.
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Abstract
backgroundCeliac disease (CD) and type 1 diabetes mellitus (DM1) are genetically associated pathologies, and it is frequent that a patient presents both. The simultaneous detection of autoimmune markers of CD and DM1 would be a rational strategy to improve their diagnosis. The aim of this work was the expression and purification of recombinant human tissue transglutaminase and the development of an immunoassay based on flow cytometry (FloCMIA multiplex) for the simultaneous and discriminative detection of autoantibodies to tissue transglutaminase (tTgA) and two of the main markers of DM1-glutamic acid decarboxylase autoantibodies (GADA) and protein tyrosine phosphatase-related IA-2 autoantibodies (IA-2A).
resultsRecombinant tTg was purified from E. coli bacteria, yielding ≈ 9.7 mg of 90-95% pure protein/L of bacterial culture. For the immunoassay, a "double paratope" model was performed and 40 serum samples of control individuals and 25 recently diagnosed patients with CD and/or DM1 were assayed. The analytical sensitivity of FloCMIA multiplex was 67% for tTgA, 76% for GADA and 71.4% for IA-2A. The specificity was 89%, 93% and 90% for tTgA, GADA and IA-2A, respectively.
conclusionsThe FloCMIA multiplex immunoassay was successfully developed as a flow cytometry-based platform for the simultaneous and discriminative screening of CD and DM1 autoantibodies. This method offers a practical, accessible, and high-throughput solution, significantly reducing costs (up to tenfold) and operational time compared to traditional assays. By utilizing a commonly available flow cytometer, detecting all immunoglobulin isotypes, and eliminating radioactive materials, FloCMIA provides a safer, more inclusive, and sensitive tool, enabling earlier therapeutic intervention and improving large-scale population screening.
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