O2 Screening of anti-tissue transglutaminase autoantibodies in pediatric patients recently diagnosed with Diabetes Mellitus, employing a novel immunoassay

Abstract

Introduction: Type 1 Diabetes Mellitus (DM1) and Celiac Disease (CD) are chronic, polygenic and multifactorial diseases associated to an immune system dysfunction. They are simultaneously presented in 10% of the patients, thus it is recommended the routine screening of CD in DM1 recently diagnosed patients. Objectives: Screening pediatric patients diagnosed with DM1 in order to evaluate the presence of anti-tissue transglutaminase antibodies (tTgA), using a method based in flow cytometry (FloCMIA: Flow Cytometric Microsphere based Immunoassay). Materials and Methods: The presence of tTgA in serum samples of 253 pediatric patients, recently diagnosed with DM1, was evaluated by FloCMIA. A "double paratope" model was used incubating the samples with polystyrene microspheres (5m) adsorbed with recombinant tissue transglutaminase (His6-tTg), ON at 4 ºC. The mixture was then incubated with biotinylated His6-tTg and the immunocomplexes formed were detected with streptavidin-Phycoerythrin and acquired in a Flow Cytometer. The results were expressed as Standard Deviation scores (SDs), considering as positive the samples with SDs > 1.7. Samples that were tTgA positive by FloCMIA were then contrasted using a commercial ELISA kit

Keywords

• screening
• diabetes
• inmunoensayo.