Protein A and protein G ELISA for the detection of IgG autoantibodies against tissue transglutaminase in childhood celiac disease.

ABSTRACT

OBJECTIVES: To investigate if the detection of celiac disease (CD) in children was improved by using alternative conjugates for assessment of tissue transglutaminase (tTG) autoantibodies. METHODS: Serum samples from 108 biopsy confirmed CD children and 42 control subjects were investigated for the presence of autoantibodies with tTG coated microplates using protein A (PA), protein G (PG), anti-IgG, or anti-IgA as conjugates. RESULTS: Of the 108 CD children, 86 (80%) were IgG-tTG positive, 91 (84%) were positive with the PA-conjugate, 94 (87%) were positive with the PG-conjugate, and 103 (95%) were IgA-tTG positive. Among the 42 controls, 4 (10%) were IgG-tTG positive, 5 (12%) were positive with both the PA- and PG conjugates, whereas 3 (7%) were IgA-tTG positive. Compared with IgG-tTG the concordance was 93% for PA and 95% for PG, with a positive correlation between antibody levels (r=0.967 and r=0.975, p<0.0001). All but one CD child were found positive by combining IgG-tTG and IgA-tTG detection. CONCLUSIONS: The sensitivity of IgG-tTG detection with ELISA increased by protein A or protein G conjugates, whereas the specificity was reduced as compared with anti-IgG conjugate. The combined measurement of IgA-tTG and IgG-tTG still seems to be the optimal procedure when screening children for CD.