Evaluating the performance of a multiparametric IgA assay for celiac disease diagnosis
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Caterina Maria Gambino
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Anna Maria Ciaccio
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.
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Research ethics: The study protocol received approval from the Ethics Committee of the University Hospital of Palermo (No. 08/2021).
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Informed consent: Informed consent was obtained from all individuals included in this study.
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Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
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Use of Large Language Models, AI and Machine Learning Tools: None.
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Conflict of interest: The authors state no conflict of interest.
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Research funding: None declared.
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Data availability: The data that support the findings of this study are available on request from the corresponding author.
References
1. Ludvigsson, JF, Leffler, DA, Bai, JC, Biagi, F, Fasano, A, Green, PH, et al.. The Oslo definitions for coeliac disease and related terms. Gut 2013;62:43–52. https://doi.org/10.1136/gutjnl-2011-301346.Search in Google Scholar PubMed PubMed Central
2. Wieser, H. Chemistry of gluten proteins. Food Microbiol 2007;24:115–19. https://doi.org/10.1016/j.fm.2006.07.004.Search in Google Scholar PubMed
3. Jabri, B, Sollid, LM. Mechanisms of disease: immunopathogenesis of celiac disease. Nat Clin Pract Gastroenterol Hepatol 2006;3:516–25. https://doi.org/10.1038/ncpgasthep0582.Search in Google Scholar PubMed
4. Asri, N, Rostami-Nejad, M, Barzegar, M, Nikzamir, A, Rezaei-Tavirani, M, Razzaghi, M, et al.. Suppressive mechanisms induced by Tregs in celiac disease. Iran Biomed J 2020;24:140–7. https://doi.org/10.29252/ibj.24.3.140.Search in Google Scholar PubMed PubMed Central
5. Gatti, S, Rubio-Tapia, A, Makharia, G, Catassi, C. Patient and community health global burden in a world with more celiac disease. Gastroenterology 2024;167:23–33. https://doi.org/10.1053/j.gastro.2024.01.035.Search in Google Scholar PubMed
6. Cappello, M, Morreale, GC, Licata, A. Elderly onset celiac disease: a narrative review. Clin Med Insights Gastroenterol 2016;9:41–9. https://doi.org/10.4137/cgast.s38454.Search in Google Scholar
7. Pancheva, R, Dolinsek, J, Panayotova, M, Yankov, I, Kofinova, D, Nikolova, S, et al.. Bridging the gap: awareness, knowledge, and challenges of living with celiac disease in Bulgaria. Nutrients 2025;17:1267. https://doi.org/10.3390/nu17071267.Search in Google Scholar PubMed PubMed Central
8. Nachman, F, Vázquez, H, González, A, Andrenacci, P, Compagni, L, Reyes, H, et al.. Gastroesophageal reflux symptoms in patients with celiac disease and the effects of a gluten-free diet. Clin Gastroenterol Hepatol 2011;9:214–9. https://doi.org/10.1016/j.cgh.2010.06.017.Search in Google Scholar PubMed
9. Kvamme, JM, Sørbye, S, Florholmen, J, Halstensen, TS. Population-based screening for celiac disease reveals that the majority of patients are undiagnosed and improve on a gluten-free diet. Sci Rep 2022;12:12647. https://doi.org/10.1038/s41598-022-16705-2.Search in Google Scholar
10. Zingone, F, Maimaris, S, Auricchio, R, Caio, GPI, Carroccio, A, Elli, L, et al.. Guidelines of the Italian societies of gastroenterology on the diagnosis and management of coeliac disease and dermatitis herpetiformis. Dig Liver Dis 2022;54:1304–19. https://doi.org/10.1016/j.dld.2022.06.023.Search in Google Scholar PubMed
11. Catassi, GN, Pulvirenti, A, Monachesi, C, Catassi, C, Lionetti, E. Diagnostic accuracy of IgA anti-transglutaminase and IgG anti-deamidated gliadin for diagnosis of celiac disease in children under two years of age: a systematic review and meta-analysis. Nutrients 2021;14:7. https://doi.org/10.3390/nu14010007.Search in Google Scholar PubMed PubMed Central
12. Aboulaghras, S, Piancatelli, D, Taghzouti, K, Balahbib, A, Alshahrani, MM, Al Awadh, AA, et al.. Meta-analysis and systematic review of HLA DQ2/DQ8 in adults with celiac disease. Int J Mol Sci 2023;24:1188. https://doi.org/10.3390/ijms24021188.Search in Google Scholar PubMed PubMed Central
13. Sheppard, AL, Elwenspoek, MMC, Scott, LJ, Corfield, V, Everitt, H, Gillett, PM, et al.. Systematic review with meta-analysis: the accuracy of serological tests to support the diagnosis of coeliac disease. Aliment Pharmacol Ther 2022;55:514–27. https://doi.org/10.1111/apt.16729.Search in Google Scholar PubMed PubMed Central
14. Bürgin-Wolff, A, Mauro, B, Faruk, H. Intestinal biopsy is not always required to diagnose celiac disease: a retrospective analysis of combined antibody tests. BMC Gastroenterol 2013;13:19. https://doi.org/10.1186/1471-230x-13-19.Search in Google Scholar
15. Shiha, MG, Nandi, N, Raju, SA, Wild, G, Cross, SS, Singh, P, et al.. Accuracy of the no-biopsy approach for the diagnosis of celiac disease in adults: a systematic review and meta-analysis. Gastroenterology 2024;166:620–30. https://doi.org/10.1053/j.gastro.2023.12.023.Search in Google Scholar PubMed
16. Ciacci, C, Bai, JC, Holmes, G, Al-Toma, A, Biagi, F, Carroccio, A, et al.. Serum anti-tissue transglutaminase IgA and prediction of duodenal villous atrophy in adults with suspected coeliac disease without IgA deficiency (Bi.A.CeD): a multicentre, prospective cohort study. Lancet Gastroenterol Hepatol 2023;8:1005–14. https://doi.org/10.1016/s2468-1253(23)00205-4.Search in Google Scholar PubMed
17. Pjetraj, D, Pulvirenti, A, Moretti, M, Gatti, S, Catassi, GN, Catassi, C, et al.. Diagnostic accuracy of IgA anti-transglutaminase assessed by chemiluminescence: a systematic review and meta-analysis. Nutrients 2024;16:2427. https://doi.org/10.3390/nu16152427.Search in Google Scholar PubMed PubMed Central
18. Lebwohl, B, Rubio-Tapia, A, Assiri, A, Newland, C, Guandalini, S. Diagnosis of celiac disease. Gastrointest Endosc Clin N Am 2012;22:661–77. https://doi.org/10.1016/j.giec.2012.07.004.Search in Google Scholar PubMed PubMed Central
19. Ramírez-Sánchez, AD, Tan, IL, Gonera-de Jong, BC, Visschedijk, MC, Jonkers, I, Withoff, S. Molecular biomarkers for celiac disease: past, present and future. Int J Mol Sci 2020;21:8528. https://doi.org/10.3390/ijms21228528.Search in Google Scholar PubMed PubMed Central
20. Catassi, C, Elli, L, Bonaz, B, Bouma, G, Carroccio, A, Castillejo, G, et al.. Diagnosis of non-celiac gluten sensitivity (NCGS): the Salerno experts’ criteria. Nutrients 2015;7:4966–77. https://doi.org/10.3390/nu7064966.Search in Google Scholar PubMed PubMed Central
21. Grossmann, K, Röber, N, Hiemann, R, Rödiger, S, Schierack, P, Reinhold, D, et al.. Simultaneous detection of celiac disease-specific IgA antibodies and total IgA. Autoimmun Highlights 2016;7:2. https://doi.org/10.1007/s13317-016-0073-2.Search in Google Scholar PubMed PubMed Central
22. Choung, RS, Khaleghi, RS, Ju, JM, Marietta, EV, Van Dyke, CT, Rajasekaran, JJ, et al.. Synthetic neoepitopes of the transglutaminase-deamidated gliadin complex as biomarkers for diagnosing and monitoring celiac disease. Gastroenterology 2019;156:582–91.e1. https://doi.org/10.1053/j.gastro.2018.10.025.Search in Google Scholar PubMed
23. Agardh, D, Matthias, T, Wusterhausen, P, Neidhöfer, S, Heller, A, Lerner, A. Antibodies against neo-epitope of microbial and human transglutaminase complexes as biomarkers of childhood celiac disease. Clin Exp Immunol 2020;199:294–302. https://doi.org/10.1111/cei.13394.Search in Google Scholar PubMed PubMed Central
24. Lerner, A, Ramesh, A, Matthias, T. Serologic diagnosis of celiac disease: new biomarkers. Gastroenterol Clin N Am 2019;48:307–17. https://doi.org/10.1016/j.gtc.2019.02.009.Search in Google Scholar PubMed
25. Husby, S, Koletzko, S, Korponay-Szabó, I, Kurppa, K, Mearin, ML, Ribes-Koninckx, C, et al.. European society paediatric gastroenterology, hepatology and nutrition guidelines for diagnosing coeliac disease 2020. J Pediatr Gastroenterol Nutr 2020;70:141–56. https://doi.org/10.1097/mpg.0000000000002497.Search in Google Scholar PubMed
26. Rubio-Tapia, A, Hill, ID, Semrad, C, Kelly, CP, Greer, KB, Limketkai, BN, et al.. American college of gastroenterology guidelines update: diagnosis and management of celiac disease. Am J Gastroenterol 2023;118:59–76. https://doi.org/10.14309/ajg.0000000000002075.Search in Google Scholar PubMed
27. Wieser, H, Soldaini, C, Ciacci, C. Non-biopsy strategy for the diagnosis of celiac disease in adults: a narrative review. Turk J Gastroenterol 2024;35:589–98. https://doi.org/10.5152/tjg.2024.24092.Search in Google Scholar PubMed PubMed Central
28. Torsten, M, Aaron, L. Microbial transglutaminase is immunogenic and potentially pathogenic in pediatric celiac disease. Front Pediatr 2018;6:389. https://doi.org/10.3389/fped.2018.00389.Search in Google Scholar PubMed PubMed Central
29. Lerner, A, Matthias, T. Microbial transglutaminase should be considered as an environmental inducer of celiac disease. World J Clin Cases 2019;7:3912–4. https://doi.org/10.12998/wjcc.v7.i22.3912.Search in Google Scholar PubMed PubMed Central
30. Spatola, BN, Kaukinen, K, Collin, P, Mäki, M, Kagnoff, MF, Daugherty, PS. Persistence of elevated deamidated gliadin peptide antibodies on a gluten-free diet indicates nonresponsive coeliac disease. Aliment Pharmacol Ther 2014;39:407–17. https://doi.org/10.1111/apt.12603.Search in Google Scholar PubMed PubMed Central
31. Reinke, Y, Zimmer, KP, Naim, HY. Toxic peptides in Frazer’s fraction interact with the actin cytoskeleton and affect the targeting and function of intestinal proteins. Exp Cell Res 2009;315:3442–52. https://doi.org/10.1016/j.yexcr.2009.06.026.Search in Google Scholar PubMed
32. Kaunisto, H, Salmi, T, Lindfors, K, Kemppainen, E. Antibody responses to transglutaminase 3 in dermatitis herpetiformis: lessons from celiac disease. Int J Mol Sci 2022;23:2910. https://doi.org/10.3390/ijms23062910.Search in Google Scholar PubMed PubMed Central
33. Vermeersch, P, Geboes, K, Mariën, G, Hoffman, I, Hiele, M, Bossuyt, X. Serological diagnosis of celiac disease: comparative analysis of different strategies. Clin Chim Acta 2012;413:1761–7. https://doi.org/10.1016/j.cca.2012.06.024.Search in Google Scholar PubMed
34. Adriaanse, M, Leffler, DA. Serum markers in the clinical management of celiac disease. Dig Dis 2015;33:236–43. https://doi.org/10.1159/000371405.Search in Google Scholar PubMed PubMed Central
35. Konopka, E, Grzywnowicz, M, Oralewska, B, Cielecka-Kuszyk, J, Trojanowska, I, Cukrowska, B. Clinical utility of quantitative multi-antibody polycheck immunoassays in the diagnosis of coeliac disease. World J Gastrointest Pharmacol Ther 2016;7:254–60. https://doi.org/10.4292/wjgpt.v7.i2.254.Search in Google Scholar PubMed PubMed Central
36. Wierdsma, NJ, van Bokhorst-de van der Schueren, MA, Berkenpas, M, Mulder, CJ, van Bodegraven, AA. Vitamin and mineral deficiencies are highly prevalent in newly diagnosed celiac disease patients. Nutrients 2013;5:3975–92. https://doi.org/10.3390/nu5103975.Search in Google Scholar PubMed PubMed Central
37. Choung, RS, Larson, SA, Khaleghi, S, Rubio-Tapia, A, Ovsyannikova, IG, King, KS, et al.. Prevalence and morbidity of undiagnosed celiac disease from a community-based study. Gastroenterology 2017;152:830–9.e5. https://doi.org/10.1053/j.gastro.2016.11.043.Search in Google Scholar PubMed PubMed Central
38. Ghunaim, M, Seedi, A, Alnuman, D, Aljohani, S, Aljuhani, N, Almourai, M, et al.. Impact of a gluten-free diet in adults with celiac disease: nutritional deficiencies and challenges. Cureus 2024;16:e74983. https://doi.org/10.7759/cureus.74983.Search in Google Scholar PubMed PubMed Central
39. Majsiak, E, Cukrowska, B, Choina, M, Bielawski, K, Cielecka-Kuszyk, J, Konopka, E, et al.. Evaluation of the usefulness of a serological test for diagnosis of celiac disease simultaneously detecting specific antibodies and total IgA. Nutrients 2022;15:202. https://doi.org/10.3390/nu15010202.Search in Google Scholar PubMed PubMed Central
40. Rozenberg, O, Lerner, A, Pacht, A, Grinberg, M, Reginashvili, D, Henig, C, et al.. A new algorithm for the diagnosis of celiac disease. Cell Mol Immunol 2011;8:146–9. https://doi.org/10.1038/cmi.2010.63.Search in Google Scholar PubMed PubMed Central
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