Anti-transglutaminase IgA ELISA: clinical potential and drawbacks in celiac disease diagnosis.
BACKGROUND
Since the identification of tissue transglutaminase (tTG) as the antigen for the anti-endomysial antibodies (EMA), several antigen-specific immunoassays have been reported for celiac disease (CD) screening. A first objective was to evaluate the suitability for CD screening of three different IgA tTG ELISAs, two of them based on guinea pig liver tTG (gp-tTG) (an in-house ELISA with a partially purified extract and a commercial ELISA with purified gp-tTG antigen) and a third recombinant human tTG (rh-tTG) ELISA. The results are compared with EMA and with the final clinical diagnosis. A second objective was to analyze antibody reactivities in those patients with anti-tTG and EMA discrepancies.


METHODS
ELISA and EMA tests were used to measure IgA anti-tTG levels in sera from 259 patients (107 had CD and 72 had Type I diabetes mellitus).


RESULTS
The purified gp-tTG ELISA was highly sensitive (97.7%) and specific (98.8%) in the detection of CD, almost equaling EMA. Rh-tTG ELISA did not improved the sensitivity of EMA, but its specificity was slightly superior. Immunoblot analysis with partially purified gp-tTG extract, the antigen most frequently used for anti-tTG detection, showed that the majority of false positives were due to IgA reactivities to contaminant proteins present in the liver antigenic extract. This low specificity was particularly problematic in diabetics.


CONCLUSIONS
Purified tTG ELISAs, either with purified guinea pig liver or recombinant human antigens, can be used as quantitative and observer-independent alternatives to the traditional and time-consuming EMA in the screening of CD.
Leon
F
F
Dept. of Immunology Hospital Ramón y Cajal, Madrid, Spain.
Camarero
C
C
R-Pena
R
R
Eiras
P
P
Sanchez
L
L
Baragaño
M
M
Lombardia
M
M
Bootello
A
A
Roy
G
G
D015153:Blotting, Western; D002446:Celiac Disease / Q000150:complications / Q000175:diagnosis* / Q000276:immunology*; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood*; D007422:Intestines / Q000473:pathology* / Q000648:ultrastructure; D008871:Microvilli / Q000473:pathology; D009130:Muscle, Smooth / Q000473:pathology; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D011503:Transglutaminases / Q000276:immunology*
D007070:Immunoglobulin A; D011503:Transglutaminases