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Related Experiment Video

Updated: Apr 18, 2026

Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein Gene Product 9.5
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Laboratory procedures update on Hirschsprung disease.

Catherine Takawira1, Stephanie D'Agostini, Suzan Shenouda

  • 1*Department of Laboratory Medicine and Pathology †Department of Pediatric Gastroenterology, Stollery Children's Hospital, University of Alberta, Edmonton, Canada ‡Department of Developmental Biology and Oncology, Faculty of Health, Medicine & Life Sciences, Maastricht University, Maastricht, The Netherlands.

Journal of Pediatric Gastroenterology and Nutrition
|January 8, 2015
PubMed
Summary

Histopathology of rectal biopsies, specifically Hematoxylin and eosin staining with or without acetylcholinesterase, remains the gold standard for diagnosing Hirschsprung disease (HD) in infants. Calretinin staining may lead to false positives and overtreatment.

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Area of Science:

  • Pediatric Pathology
  • Gastroenterology
  • Surgical Pathology

Background:

  • Hirschsprung disease (HD) diagnosis relies on detecting ganglion cells in rectal biopsies.
  • Early diagnosis of HD is crucial to prevent severe complications like enterocolitis.
  • Various diagnostic techniques have been evaluated, including imaging, manometry, and histology.

Purpose of the Study:

  • To systematically review and update laboratory procedures for detecting ganglion cells in infant rectal biopsies.
  • To evaluate the diagnostic accuracy of different methods for Hirschsprung disease.

Main Methods:

  • Systematic review and meta-analysis following PRISMA guidelines.
  • Calculation of sensitivity and specificity for various diagnostic tests.
  • Review of immunohistochemical markers, including calretinin.

Main Results:

  • Contrast enema demonstrated the lowest sensitivity and specificity.
  • Biopsy with histology showed the highest sensitivity and specificity.
  • Hematoxylin and eosin (H&E) staining, with or without acetylcholinesterase, is highly accurate; calretinin staining showed potential for false positives.

Conclusions:

  • H&E staining, with or without acetylcholinesterase, is the criterion standard for HD diagnosis.
  • Calretinin staining's high false-positive rate may limit its use and lead to overtreatment.
  • Diagnostic strategies should balance accuracy with the principle of 'primum non nocere' to avoid unnecessary surgeries.