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Aim: Hirschsprung’s disease (HD) is the congenital aganglionosis of the human colon due to disruptionof migration, imbalanced proliferation and differentiation of neural crest cell. The diagnosis and extent ofresection for the management of HD depend on the sensitive and specific identification of ganglion cells.Negative immunohistochemical expression of calretinin, shown its superiority as diagnostic method for fullthickness as well as partial thickness rectal suction biopsies.Materials and Methods: Total 86 colonic and rectal biopsies of 43 patients attending Sassoon hospital,Pune (between 2015-2017) with surgically resected specimen proven Hirschsprung’s disease cases andfollow up cases of non HD cases were evaluated with both Haematoxylin and Eosin stain and calretininIHC.Result: Age group was 2 days to 7 year with having Male: Female ratio 2.91:1. Out of 43 cases, 26 werediagnosed as Hirschsprung’s Disease, 16 were not having Hirschsprung’s disease, 1 case of hypogangliosis.13 out of 86 biopsy and 9 out of 43 cases show false positive result with use of Haematoxylin and Eosinstain only. Specificity and negative predictive value of H&E stain was 72% and 78.12% respectively withaccuracy of 86%. Use of calretinin IHC gave 100% accuracy in all 86 biopsies studied including 17 partialthickness biopsies.Conclusion: Only use of Haematoxylin and Eosin stain will lead to significant false positive result leadingto misdiagnosis and in turn wrong treatment. Loss of calretinin expression in aganglionic segment help inthe interpretation of even partial biopsies and easy to interpret with improvement in diagnosis upto 100%.Keywords: Hirschsprung’s disease, Calretinin, Rectal biopsy.

To assess calretinin immunohistochemistry in Hirschsprung’s disease and allied disorders