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Evaluating the performance of a multiparametric IgA assay for celiac disease diagnosis

  • Caterina Maria Gambino , Luisa Agnello , Fabio Del Ben ORCID logo , Anna Maria Ciaccio , Salvatore Milano , Roberta Vassallo , Francesco Cacciabaudo , Aurelio Seidita , Pasquale Mansueto , Antonio Carroccio and Marcello Ciaccio EMAIL logo
Published/Copyright: August 20, 2025
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 63 Issue 12

Abstract

Objectives

Celiac disease (CD) is a systemic autoimmune disorder triggered by gluten in genetically predisposed individuals. Accurate diagnosis remains challenging due to clinical heterogeneity and reliance on invasive biopsy. This study aimed to evaluate the diagnostic performance of a novel multiparametric membrane-based enzyme immunoassay (AESKUBLOTS®) for the simultaneous detection of IgA antibodies targeting eight CD-related antigens.

Methods

A retrospective, single-centre study was conducted on 180 participants: 80 with CD (30 untreated, 50 on gluten-free diet, GFD), 50 with non-celiac wheat sensitivity (NCWS), and 50 healthy controls (HC). Serum samples were analysed using the AESKU assay. Diagnostic accuracy was assessed via ROC curve analysis and 5-fold cross-validation, examining individual markers and a composite antibody score.

Results

The assay demonstrated high diagnostic performance, particularly in untreated CD patients. Anti-tTG neo IgA showed the highest accuracy (AUC=0.93), followed by anti-tTG IgA (AUC=0.92). A composite score of ≥4 positive markers yielded an AUC of 0.99, while ≥6 positive markers achieved 100 % specificity and PPV, with 76.7 % sensitivity. Notably, anti-mTG IgA levels were elevated in all CD patients regardless of diet, suggesting potential utility in monitoring or identifying ongoing mucosal immune activity.

Conclusions

This multiparametric IgA assay offers a sensitive, specific, and non-invasive diagnostic tool for CD. Larger, prospective studies are warranted to confirm the clinical utility and expand the applicability to broader populations.

Keywords: autoimmunity; microbial transglutaminase; multiparametric test; neo-epitopes
Corresponding author: Prof. Marcello Ciaccio, Department of Biomedicine, Neurosciences and Advanced Diagnostics, Institute of Clinical Biochemistry, Clinical Molecular Medicine, and Clinical Laboratory Medicine, University of Palermo, Palermo, Italy; and Department of Laboratory Medicine, University Hospital Paolo Giaccone, Palermo, Italy, E-mail:
Caterina Maria Gambino and Luisa Agnello contributed equally to this work as first authors.

  1. Research ethics: The study protocol received approval from the Ethics Committee of the University Hospital of Palermo (No. 08/2021).

  2. Informed consent: Informed consent was obtained from all individuals included in this study.

  3. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  4. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  5. Conflict of interest: The authors state no conflict of interest.

  6. Research funding: None declared.

  7. Data availability: The data that support the findings of this study are available on request from the corresponding author.

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Received: 2025-06-09
Accepted: 2025-08-07
Published Online: 2025-08-20
Published in Print: 2025-11-25

© 2025 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2025-0705
Keywords for this article
autoimmunity; microbial transglutaminase; multiparametric test; neo-epitopes

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. Challenging the dogma: why reviewers should be allowed to use AI tools
  4. Multivariate approaches to improve the interpretation of laboratory data
  5. Review
  6. Interference of therapeutic monoclonal antibodies with electrophoresis and immunofixation of serum proteins: state of knowledge and systematic review
  7. Opinion Papers
  8. Urgent call to the European Commission to simplify and contextualize IVDR Article 5.5 for tailored and precision diagnostics
  9. The importance of laboratory medicine in the management of CKD-MBD: insights from the KDIGO 2023 controversies conference
  10. Supplementation of pyridoxal-5′-phosphate in aminotransferase reagents: a matter of patient safety
  11. HCV serology: an unfinished agenda
  12. From metabolic profiles to clinical interpretation: multivariate approaches to population-based and personalized reference intervals and reference change values
  13. Genetics and Molecular Diagnostics
  14. A multiplex allele specific PCR capillary electrophoresis (mASPCR-CE) assay for simultaneously analysis of SMN1/SMN2/NAIP copy number and SMN1 loss-of-function variants
  15. General Clinical Chemistry and Laboratory Medicine
  16. From assessment to action: experience from a quality improvement initiative integrating indicator evaluation and adverse event analysis in a clinical laboratory
  17. Evaluation of measurement uncertainty of 11 serum proteins measured by immunoturbidimetric methods according to ISO/TS 20914: a 1-year laboratory data analysis
  18. Assessing the harmonization of current total vitamin B12 measurement methods: relevance and implications
  19. The current status of serum insulin measurements and the need for standardization
  20. Method comparison of plasma and CSF GFAP immunoassays across multiple platforms
  21. Cerebrospinal fluid leptin in Alzheimer’s disease: relationship to plasma levels and to cerebrospinal amyloid
  22. Verification of the T50 Calciprotein Crystallization test: bias estimation and interferences
  23. An innovative immunoassay for accurate aldosterone quantification: overcoming low-level inaccuracy and renal dysfunction-associated interference
  24. Oral salt loading combined with postural stimulation tests for confirming and subtyping primary aldosteronism
  25. Evaluating the performance of a multiparametric IgA assay for celiac disease diagnosis
  26. Clinical significance of anti-mitochondrial antibodies and PBC-specific anti-nuclear antibodies in evaluating atypical primary biliary cholangitis with normal alkaline phosphatase levels
  27. Reference Values and Biological Variations
  28. Establishment of region-, age- and sex-specific reference intervals for aldosterone and renin with sandwich chemiluminescence immunoassays
  29. Validation of a plasma GFAP immunoassay and establishment of age-related reference values: bridging analytical performance and routine implementation
  30. Comparative analysis of population-based and personalized reference intervals for biochemical markers in peri-menopausal women: population from the PALM cohort study
  31. Hematology and Coagulation
  32. Evaluation of stability and potential interference on the α-thalassaemia early eluting peak and immunochromatographic strip test for α-thalassaemia --SEA carrier screening
  33. Cardiovascular Diseases
  34. Analytical and clinical evaluation of an automated high-sensitivity cardiac troponin I assay for whole blood
  35. Diabetes
  36. Method comparison of diabetes mellitus associated autoantibodies in serum specimens
  37. Letters to the Editor
  38. Permitting disclosed AI assistance in peer review: parity, confidentiality, and recognition
  39. Response to the editorial by Karl Lackner
  40. Hemolysis detection using the GEM 7000 at the point of care in a pediatric hospital setting: does it affect outcomes?
  41. Estimation of measurement uncertainty for free drug concentrations using ultrafiltration
  42. Cryoglobulin pre-analysis over the weekend
  43. Accelerating time from result to clinical action: impact of an automated critical results reporting system
  44. Recent decline in patient serum folate test levels using Roche Diagnostics Folate III assay
  45. Kidney stones consisting of 1-methyluric acid
  46. Congress Abstracts
  47. 7th EFLM Conference on Preanalytical Phase
  48. Association of Clinical Biochemists in Ireland Annual Conference
  49. Association of Clinical Biochemists in Ireland Annual Conference
  50. 17th Congress of the Portuguese Society of Clinical Chemistry, Genetics and Laboratory Medicine
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