Abstract
Celiac disease (CD) is diagnosed by a combination of specific serology and typical duodenal lesions. The histological confirmation of CD, mandatory in the majority of patients with suspected CD, is based on invasive and poorly tolerated procedures, such as upper gastrointestinal endoscopy. In this study we propose an alternative and non-invasive methodology able to confirm the diagnosis of CD based on the analysis of serum samples using the Raman spectroscopy technique. Three different bands centered at 1650, 1450 and 1003 cm−1 have been considered and the A1450/A1003 and A1650/A1003 ratios have been computed to discriminate between CD and non-CD subjects. The reliability of the methodology was validated by statistical analysis using receiver operating characteristic (ROC) curves. The Youden index was also determined to obtain optimal cut-off points. The obtained results highlighted that the proposed methodology was able to distinguish between CD and non-CD subjects with 98% accuracy. The optimal cut-off points revealed, for both the A1450/A1003 and A1650/A1003 ratios, high values of sensitivity and specificity (>95.0% and >92.0% respectively), confirming that Raman spectroscopy may be considered a valid alternative to duodenal biopsy and demonstrates spectral changes in the secondary structures of the protein network.
Highlights
Celiac disease (CD) is an immuno-mediated systemic disorder due to ingestion of gluten proteins of wheat, barley, and rye in genetically susceptible individuals carrying the HLA-DQ2 and/or -DQ8 alleles [1]
Given the crucial role of transglutaminase in CD, it is not surprising that the spectrum of CD patients differs from that of controls in specific regions involving conformational modification of proteins concerned in transamidation and deamidation by transglutaminase. tTG is expressed in small intestine, and that expression is increased in untreated celiac small intestinal mucosa, where tTG is detected at the level of muscularis mucosae and pericryptal fibroblasts adjacent to enterocytes [56]
Cell surface tTG was found on macrophages and dendritic cells, which are known to play an important role in the pathogenesis of CD. tTG expression was detected in celiac enterocytes, together with an evident upregulation of the enzymatic activity in the subepithelial layer
Summary
Celiac disease (CD) is an immuno-mediated systemic disorder due to ingestion of gluten proteins of wheat, barley, and rye in genetically susceptible individuals carrying the HLA-DQ2 and/or -DQ8 alleles [1]. A combination of CD serology testing and duodenal biopsy sampling is required for the diagnosis of CD in adults [4], but not in all children, provided that they meet the criteria suggested by the new ESPGHAN guidelines [5]. These criteria rely on measurement of the concentration of tTG (anti-tTG) IgA antibodies (TGA-IgA). [36,37] Raman spectroscopy was used to reveal biochemical differences in the plasma of Crohn’s disease patients and to carry out Celiac disease diagnosis on individual red blood cells, respectively. In this last study [37], chemometric analysis were used in combination with Raman hyperspectroscopy, because insufficient differentiation between average Raman spectra of Celiac disease donors and healthy patients was evident
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