A modified ELISA for improved detection of IgA, IgG, and IgM anti-tissue transglutaminase antibodies in celiac disease

Clin Chim Acta. 2009 May;403(1-2):37-41. doi: 10.1016/j.cca.2009.01.006.

Abstract

Background: Serum IgA antibodies against tissue transglutaminase (tTG) are reliable markers for celiac disease (CD), still the diagnostic performance of anti-tTG immunoassays can be improved. A novel ELISA, using fibronectin (FN) as tTG binding protein, was evaluated for the detection of anti-tTG antibodies in CD.

Methods: Sera from 173 children with untreated CD and 97 controls were analyzed for IgA, IgG, and IgM anti-tTG antibodies with ELISA using human recombinant tTG with or without FN.

Results: The areas under the ROC (receiver operating characteristic) curves were significantly higher for FN/tTG ELISA compared to tTG ELISA for IgG (0.930 versus 0.809; p < 0.001) and IgM (0.850 versus 0.811; p = 0.019), but not for IgA (0.977 versus 0.970; p = 0.356), respectively. At the fixed diagnostic specificity (100% for IgA and IgM, 99% for IgG), the sensitivity of all three FN/tTG ELISA (92.5% for IgA, 60.7% for IgG, 50.3% for IgM) exceeded those obtained in tTG ELISA (89.0% for IgA, 48.6% for IgG, 38.7% for IgM; p < 0.05). The combined use of IgA- and IgG-FN/tTG ELISA resulted in 95.4% sensitivity and 99.0% specificity for CD.

Conclusions: Using FN to bind tTG improves the diagnostic performance of solid phase anti-tTG antibody assays for childhood CD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Chemical Analysis / methods*
  • Celiac Disease / blood
  • Celiac Disease / diagnosis*
  • Celiac Disease / immunology*
  • Child
  • Child, Preschool
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Fibronectins / metabolism
  • Humans
  • Immunoglobulin A / blood
  • Immunoglobulin A / immunology
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology
  • Immunoglobulin Isotypes / blood*
  • Immunoglobulin Isotypes / immunology*
  • Immunoglobulin M / blood
  • Immunoglobulin M / immunology
  • Male
  • Transglutaminases / immunology*
  • Transglutaminases / metabolism

Substances

  • Fibronectins
  • Immunoglobulin A
  • Immunoglobulin G
  • Immunoglobulin Isotypes
  • Immunoglobulin M
  • Transglutaminases