Flow cytometric detection of anti-gliadin antibodies

Abstract

A very sensitive solid-phase fluorescent immunoassay to detect anti-α-gliadin IgA class antibodies is described. The solid phase consisted of polystyrene carboxylated microspheres, of 5 μm diameter, coated with α-gliadin. Serum-specific antibodies bound to the α-gliadin were measured by flow cytometry using fluorescein-conjugated anti-human IgA. 41 samples were tested and the results compared with those obtained by a standard method: an enzyme-linked immunosorbent assay (ELISA). A good correlation was found between the two techniques ( r = 0.96). The sera of untreated coeliac children showed significantly higher antibody values than the sera of children on a gluten-free diet or healthy control groups. The flow cytometric method was more sensitive when the Kolgomorov/Smirnov test was used to analyse the histograms. This method provides an alternative screening test for coeliac disease and may also be used to confirm borderline results obtained in the ELISA test.