Abstract
Objective: Development of high performance serological tests for diagnosis of celiac disease. ELISA tests using the calcium dependent, cross linking enzyme tissue transglutaminase (tTG) have proven to be useful for aiding in serological diagnosis of celiac disease and related gluten sensitive enteropathies. Sensitivity of these newer IgA ELISA tests is compromised due to 1–3% of celiac patients being IgA deficient. Specificity can be compromised by use of low specific activity antigen preparations.Methods: ELISA plates are coated with high specific activity tissue transglutaminase isolated from fresh human red blood cells. Anti-human IgA and IgG HRP labeled conjugates and a common diluent, wash and chromogen are used to test 185 normal samples, 90 patients with related gastrointestinal or connective tissue disease and a group of biopsy proven IgA sufficient and deficient celiacs.Results: All 24 active, IgA sufficient celiac patients were found to be IgA tTG positive as were 10 of 11 pediatric patients. As expected, all 16 IgA deficient celiac patients were negative by traditional IgA endomysial and tTG ELISA assays. Fifteen of these 16 patients were found to be positive for IgG antibodies to tTG. The one negative sample was already on diet therapy. Specificity for both the IgA and IgG versions of the human red blood cell tTG test was 99%.The ELISA incorporating human red blood cell tTG was somewhat more sensitive and specific compared with first generation tTG Elisas using calcium optimized antigen purified from guinea pig liver. When compared against several recombinantly derived human tTG antigens, the native human red blood cell antigen exhibited higher signal to noise ratio and proved to be easier and more reliably purified.Conclusion: Detection of IgA and IgG autoantibodies to high specific activity human red blood cell derived tTG affords a sensitive and specific serologic system for aiding in diagnosis of celiac disease. The ELISAs are objective, automatable and cost effective alternatives to standard immunofluorescence based endomysial antibody tests.
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