# Histopathologic Concerns and Diagnostic Challenges in Hirschsprung’s Disease: An Eastern European Single-Center Observational Study

## TL;DR

This study examines diagnostic challenges in Hirschsprung’s disease using immunohistochemistry and morphometry in biopsy and surgical specimens.

## Contribution

The study introduces a coupled calretinin–CD56 immunohistochemistry approach to improve diagnosis in challenging cases.

## Key findings

- Calretinin–CD56 immunohistochemistry helps diagnose ganglion cell paucity in biopsies with incomplete submucosa.
- Re-biopsy is recommended when calretinin-positive nerve fibrils are present without nerve trunk hypertrophy.
- Nerve trunk size differs significantly between biopsies and surgical specimens, emphasizing the need for consistent assessment.

## Abstract

Background: We proposed a comprehensive clinicopathological study involving the characterization of the study cohort and a comparative analysis of biopsies and surgical specimens from patients with Hirschsprung’s disease. The study was complemented by the diagnostic value of calretinin, CD56, and S-100 immunohistochemistry. Methods: Descriptive statistical analysis of diagnostic variables in the group of biopsy specimens (n = 32) and bowel resection specimens (n = 16) was performed. The pattern of calretinin and CD56 expression in Meissner’s plexus elements was analyzed and the maximum thicknesses of the nerve fibers were measured using morphometry with S100-immunostained sections. Conclusions: Coupled calretinin–CD56 immunohistochemistry is useful in diagnosing ganglion cell paucity biopsies or specimens with incomplete submucosa. In cases where there are no ganglion cells but there are calretinin-positive nerve fibrils in the lamina propria without nerve trunk (NT) hypertrophy, re-biopsy is the best solution. The significant differences in NT size between biopsies and surgical specimens highlight the importance of assessing NT diameter in all tissue samples examined.