A beta-turn rich oats peptide as an antigen in an ELISA method for the screening of coeliac disease in a paediatric population.
BACKGROUND
ELISA methods for the measurement of IgA antigliadin antibodies (AGA), both home-made and commercial systems, routinely employ wheat gliadin fractions as coating antigens. We investigate the sensitivity and specificity for CD diagnosis of a new ELISA method using a highly immunoreactive beta-turn rich gamma3-avenin peptide as an alternative coating antigen.


METHODS
The assay was standardized with antihuman IgA peroxidase-conjugated as the second antibody. Alternatively, an ELISA based on the use of protein A-peroxidase was assayed to measure both IgG plus IgA antibodies. Sixty-three sera from healthy controls were analyzed to establish the system's cut-off point. Sera from 103 coeliac and from 65 noncoeliac children were tested; for diagnosis purposes, a small intestinal biopsy had been performed in all of them.


RESULTS
For the IgA class antibodies assay a high sensitivity and specificity of 90.3% and 98.5%, respectively, was obtained, comparable to those achieved for IgA antiendomysium antibodies (EmA) with the same sera.


CONCLUSIONS
In view of the high sensitivity and specificity obtained together with water solubility of the peptide and easiness for large-scale reproducible synthesis, the new AGA IgA avenin peptide ELISA represents a significant improvement in CD diagnosis in comparison with conventional established AGA IgA ELISA using crude gliadins as coating antigens.
Ribes-Koninckx
C
C
Hospital La Fe, Valencia, Spain.
Alfonso
P
P
Ortigosa
L
L
Escobar
H
H
Suárez
L
L
Arranz
E
E
Méndez
E
E
D000293:Adolescent; D000328:Adult; D000906:Antibodies / Q000097:blood*; D018554:Avena / Q000737:chemistry / Q000276:immunology*; D002446:Celiac Disease / Q000175:diagnosis* / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005903:Gliadin / Q000276:immunology; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D008297:Male; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology*; D011237:Predictive Value of Tests; D055315:Prolamins; D017433:Protein Structure, Secondary; D012680:Sensitivity and Specificity
D000906:Antibodies; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D010455:Peptides; D010940:Plant Proteins; D055315:Prolamins; D005903:Gliadin