Frequency of loss of calretinin expression in clinically susceptible cases of Hirschprung disease in rectal biopsies


  • Ayesha Haider Combined Military Hospital, Multan Pakistan
  • Unaiza Jamil Combined Military Hospital, Multan Pakistan
  • Iqra Ahmad Shah Combined Military Hospital, Multan Pakistan
  • Maria Aslam Combined Military Hospital, Abbottabad Pakistan
  • Kiran Mumtaz Combined Military Hospital, Multan Pakistan
  • Syed Naeem Raza Hamdani Combined Military Hospital, Multan Pakistan



Objective: To determine the frequency of loss of calretinin expression in clinically susceptible cases of Hirschprung disease in rectal biopsies.

Material and Methods: This descriptive, cross-sectional study was conducted at Department of Histopathology, Combined Military Hospital, Multan for a period of 6 months (17th Oct 2021 to 16th April 2022). A total 93 suspected cases of Hirschprung disease were taken. Specimens received in 10 % buffered formalin and fulfilling inclusion criteria were included in the study. The cases diagnosed as Hirschprung disease were taken and calretinin immunohistochemical stain was applied and result recorded after evaluation by consultant histopathologist. Data was analyzed using (Statistical Software for Social Sciences (SPSS) version 19).

Results: Out of 93 cases, 67 (72.0%) were male patients, whereas 26 (28.0%) were female patients. The average age of the study participants was 7.95 months, with a minimum age of 1 month and a maximum age of 12 months. Twenty-eight (30.1%) of the 93 study cases were from rural areas, while 65 (69.9%) were from metropolitan areas. Twenty-five families (26.9%) reported monthly incomes of up to Rs. 35000, while 68 families (73.1%) reported incomes of over Rs. 35,000. Of these 93 study cases, failure to pass meconium was noted in 24 (25.8%) patients, constipation was noted in 75 (80.6%) and abdominal distension was noted in 62 (66.7%) patients. Loss of calretinin expression was noted in 34 (36.6%) of our study cases. 

Conclusion: According to the findings of our investigation, calretinin is very helpful in identifying suspicious cases of Hirschsprung disease. When acetylcholinesterase enzyme histochemistry is not accessible, it can be a helpful, cost-effective diagnostic assistance in centers with limited resources. It is a reliable and efficient diagnostic tool for early diagnosis of the illness. Loss of Calretinin expression was significantly associated with age and residential status.

Keywords: Calretinin expression, Frequency, Hirschsprung disease, Immunohistochemistry


Butler RT, Jaden NE, Trainor PA. The developmental etiology and pathogenesis of Hirschsprung disease. Transl Res. 2013; 162(1): 1-15. DOI:

Rosai J. Hirschprung’s disease and related disorders. In Rosai J, Ackerman L, editors. Rosai and Ackerman’s surgical pathology.10th ed. St .Louis: Mosby; 2011.p.732-34

Hiradfar M, Sharifi N, Khajedaluee M, Zabolinejad N, Jamshidi ST. Calretinin immunohistochemistry: An aid in the diagnosis of Hirschprung’s disease. Iran J Basic Med Sci. 2012; 15(5): 1053-9.

Alexandrescu S, Rosenberg H, Tatevian N. Role of calretinin immunohistochemical stain in evaluation of Hirschsprung disease: An institutional experience. Int J clin Exp Pathol. 2013; 6(12): 2955-61.

Kapur RP, Reed RC, Finn LS, Patterso K, Johanson J, Rutledge JC. Calretinin immunohistochemistry versus acetyl-cholinesterase histochemistry in the evaluation of suction rectal biopsies for Hirschprung disease. Pediatr Dev Pathol. 2009; 12(1): 6-15. DOI:

Samuel VG, Bonnard A, Lagausie PD, Philippe-Chomette P, Albert C, Ghoneimi AE, et al. Calretinin immunohistochemistry: A simple and efficient tool to diagnose Hirschprung disease. Mod Pathol. 2009; 22: 1379–84


Kannaiyan L, Madabhushi S, Malleboyina R, Are NK, Reddy KR, Rao B. Calretinin immunohistochemistry: A new cost-effective and easy method for diagnosis of Hirschprung’s disease. J Indian Assoc Pediatr Surg. 2013; 18(2): 66–8.


van den Berg MM, Benninga MA, Di Lorenzo C. Epidemiology of childhood constipation: A systematic review. Am J Gastroenterol. 2006; 101: 2401-9. DOI:

Rubin GP. Childhood constipation. Am Fam Physician. 2003; 67: 1041-2.

Di Lorenzo C. Pediatric anorectal disorders. Gastroenterol Clin North Am. 2001; 30: 26987.

Constipation Guideline Committee of the North American Society for Pediatric Gastroenterology, Hepatology and Nutrition. Evaluation and treatment of constipation in infants and children: Recommendations of the North American Society for Pediatric Gastroenterology, Hepatology and Nutrition. J Pediatr Gastroenterol Nutr. 2006; 43: e113.

Hyman PE, Milla PJ, Benninga MA, Davidson GP, Fleisher DF, Taminiau J. Childhood functional gastrointestinal disorders: Neonate/ toddler. Gastroenterology. 2006; 130: 1519-26. DOI:

Benninga MA, Büller HA, Heymans HS, Tytgat GN, Taminiau JA. Is encopresis always the result of constipation? Arch Dis Child. 1994; 71: 186-93.

Thompson WG, Longstreth GF, Drossman DA, Heaton KW, Irvine EJ, Müller-Lissner SA. Functional bowel disorders and functional abdominal pain. Gut. 1999; 45 Suppl 2: II43-7. DOI:

Partin JC, Hamill SK, Fischel JE, Partin JS. Painful defecation and fecal soiling in children. Pediatrics. 1992; 89: 1007-9.

Di Lorenzo C. Childhood constipation: Finally some hard data about hard stools! J Pediatr. 2000; 136: 4-7. DOI:

Abrahamian FP, Lloyd-Still JD. Chronic constipation in childhood: A longitudinal study of 186 patients. J Pediatr Gastroenterol Nutr. 1984; 3: 460-7.

Dingemans AJ, van der Steeg HJ, Rassouli-Kirchmeier R, Linssen MW, van Rooij IA, de Blaauw I. Redo pull-through surgery in Hirschsprung disease: short-term clinical outcome. J Pediatr Surg. 2017;52(9):1446-50. DOI:

Khan K. Two stage management of patients with Hirschsprung`s disease. J Surg Pak Int. 2005; 10(4): 7-10.

Henna N, Sheikh A, Shaukat M, Nagi AH. Children with clinical presentation of hirschsprung's disease - a clinicopathological experience. Biomedica. 2011; 27(1): 1-4.

Hussain S, Mazhar A, Siddiqui A. Transanal endorectal pull-through: An effective alternative to staged management of Hirschsprung’s disease. Pak J Surg. 2013; 29(3): 17982.

Zamir N, Akhtar J. Hirschsprung`s disease: Pattern of clinical presentation. J Surg Pak (Int). 2014; 19(1): 40-4.




How to Cite

Haider A, Jamil U, Shah IA, Aslam M, Mumtaz K, Hamdani SNR. Frequency of loss of calretinin expression in clinically susceptible cases of Hirschprung disease in rectal biopsies. Pak J Pathol [Internet]. 2024 Mar. 29 [cited 2024 Apr. 23];35(1):20-5. Available from: