Hirschsprung's disease (HD) is the most common cause of colon obstruction in neonates. Lack of calretinin immunohistochemistry (IHC) in colon lamina propria could be associated with aganglionosis. Methods: This cross-sectional retrospective study examined 64 patients suspicious of HD and conducted on collected-demographic data and Hematoxylin/Eosin (H & E) results of the surgical rectal biopsy from the archive of Imam Hossein Hospital, Isfahan, Iran. Calretinin IHC was achieved for all the

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... mens, and the results of the two methods were compared. The receiver operating characteristic (ROC) chart shows sensitivity changes according to (1-specificity). Moreover, sensitivity, specificity, as well as the positive and negative predictive values (PPV and NPV) of calretinin staining in lamina propria and muscularis mucosa, compared to H & E staining, was calculated in this study. Results: The number of males and females was the same in this study (n=32; 50%). The age of the patients ranged from 1 month to 156 months (13 years). Out of 64 patients, 8 cases were diagnosed with HD, and calretinin staining was negative in 7 patients; moreover, only one case showed weak positive staining. In 56 patients, who were not HD, two patients with negative calretinin staining had a positive H&E staining report. The current study aimed to check the diagnostic value of calretinin IHC in lamina propria of patients suspicious of HD. Compared to H&E staining, which is the routine method in thick wall biopsies, the statistics show that calretinin IHC has a high diagnostic value (P<0.001), sensitivity of 87%, specificity of 96%, PPV of 77%, and NPV of 98%. Conclusion: In conclusion, calretinin IHC utilized in specimens seems superficial and inadequate; moreover, the association with the high specificity percentage (96%) could help physicians with less invasive biopsy methods, such as endoscopic rectal biopsy.