title: Validation of a new enzyme-linked immunosorbent assay to detect the triggering proteins and peptides for celiac disease: interlaboratory study.
abstract: The performance of Gluten-Tec (EuroProxima, Arnhem, The Netherlands) was tested through an interlaboratory study in accordance with AOAC guidelines. Gluten-Tec is a competitive ELISA that detects an immunostimulatory epitope of a-gliadin in dietary food for celiacs. Fifteen laboratories, representing 14 different countries, announced their interest in taking part in this study. Of the 12 laboratories that sent the results within the established timeframe, two submitted inappropriate standard curves and were excluded from the statistical analysis. Four different food matrixes (rice-based baby food, maize bread, chocolate cake mix, and beer) were selected for preparing the test samples. Two gliadin extraction procedures were used: the conventional 60% ethanol, and a new method based on the reducing reagent dithiothreitol. The 38 samples (19 blind duplicates) tested in this study were prepared by diluting the different extracts in order to cover a wide range of gliadin levels. Both sample extraction and dilution were performed by EuroProxima; the present interlaboratory study was focused only on testing the ELISA part of the Gluten-Tec kit protocol. Repeatability values (within-laboratory variance), expressed as RSD(r) ranged from 6.2 to 25.7%, while reproducibility values (interlaboratory variance), expressed as RSD(R), ranged from 10.6 to 45.9%. Both statistical parameters were in the acceptable range of ELISAs under these conditions, and the method will be presented to the Codex Alimentarius as a preferred method for gluten analysis.
authors.affiliation: Leiden University Medical Center, Department of Immunology and Blood Transfusion, Leiden, The Netherlands. jrmujico@gmail.com
authors.forename: Jorge R
authors.initials: JR
authors.lastname: Mujico
authors.forename: Liesbeth
authors.initials: L
authors.lastname: Dekking
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Kooy-Winkelaar
authors.forename: Ron
authors.initials: R
authors.lastname: Verheijen
authors.forename: Piet
authors.initials: P
authors.lastname: van Wichen
authors.forename: Lucia
authors.initials: L
authors.lastname: Streppel
authors.forename: Nermin
authors.initials: N
authors.lastname: Sajic
authors.forename: Jan W
authors.initials: JW
authors.lastname: Drijfhout
authors.forename: Frits
authors.initials: F
authors.lastname: Koning
mesh_terms: D000485:Allergens / Q000032:analysis; D001515:Beer / Q000032:analysis; D002446:Celiac Disease / Q000139:chemically induced*; D002851:Chromatography, High Pressure Liquid; D004044:Dietary Proteins / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005504:Food Analysis; D005512:Food Hypersensitivity / Q000276:immunology; D005903:Gliadin / Q000032:analysis / Q000302:isolation & purification; D005983:Glutens / Q000032:analysis; D006801:Humans; D007202:Indicators and Reagents; D007223:Infant; D007225:Infant Food; D057230:Limit of Detection; D010455:Peptides / Q000032:analysis*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results
chemical_list: D000485:Allergens; D004044:Dietary Proteins; D007202:Indicators and Reagents; D010455:Peptides; D011933:Reagent Kits, Diagnostic; D005983:Glutens; D005903:Gliadin