MULTIPLEX IMMUNOASSAY FOR SIMULTANEOUS DIAGNOSTIC OF AUTOIMMUNE DIABETES MELLITUS AND CELIAC DISEASE

Abstract

Introduction: autoimmune diabetes mellitus (ADM) and celiac disease (CD) are chronic, polygenic and multifactorial diseases associated with immune system dysfunction. As it is frequent that a patient presents both pathologies, the simultaneous detection of autoimmunity markers of ADM and CD would be a rational strategy to improve the diagnosis. Objectives: to develop an immunoassay based on Flow Cytometry (FloCMIA multiplex) for the simultaneous and discriminative detection of markers for ADM (GADA and IA-2A) and CD (tTgA). Materials and methods: thirty five serum samples of control individuals and 21 type 1 DM patients were assayed. A “double bridge” model was used for the assay, incubating the serum samples with a mixture of microspheres containing different amount of internal fluorescence, each one adsorbed with an autoantigen: TrxGAD, TrxIA-2 or H6tTg, and a mixture of the same biotinilated autoantigens. The immunocomplexes were detected using streptavidin-phycoerytrin and then acquired in a flow cytometer. Conclusions: FloCMIA multiplex allowed detecting and discriminating GADA, IA-2A and/or tTgA, -in a single assay- in serum samples of ADM and/or CD. The novel developed immunoassay simplifies the screening of the large scale population.

Keywords

• diabetes mellitus autoinmune
• enfermedad celíaca
• autoanticuerpos
• inmunoensayo multiplex.