Calretinin Immunohistochemistery: An Aid in the Diagnosis of Hirschsprung’s Disease

Document Type : Original Article

Authors

1 Department of Pediatric Surgery, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Pathology, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Community Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s)
Definite diagnosis of Hirschsprung’s disease (HD) is based on histopathological study, but there are limitations associated with standard histology and histochemistry in this regard. The aim of this study was to investigate calretinin immunostaining patterns in both ganglionic and aganglionic HD intestinal specimens and to compare them with control specimens.
Materials and Methods
Specimens included 30 patients with histopathologic diagnosis of HD and 20 patients that underwent colectomy for other reasons (as control group). Eighty paraffin wax blocks of full thickness intestinal specimens (30 blocks of ganglionic segments, 30 blocks of aganglionic segments and 20 blocks of control group) were studied. Calretinin immunoreactivity and pattern of staining for ganglion cells (nuclear and cytoplasmic) and also nerve fibers in different layers of bowel were evaluated in IHC stained slides.
Results
There were positive immunostaining of nerve fibers in the lamina propria, submucosa and muscularis propria in control and patient group. There were also nuclear and cytoplasmic staining of ganglion cells in submucosa and muscularis propria in all specimens of both control group (100%) and ganglionic segments (100%). Calretinin immunoexpression of nerve fibers in muscularis propria of the aganglionic segments was negative in all but two cases (6.7%). This method had sensitivity of 93.3% and specificity of 100% for diagnosis of HD in full thickness specimens of intestinal wall. The positive predictive value was 100% and negative predictive value was 93.8%.
Conclusion
Calretinin immunohistochemistry can be used on suction rectal biopsies as a reliable and adjunctive method to diagnose HD.

Keywords

References

1. Haricharan RN, Georgeson KE. Hirschsprung disease. Semin Pediatr Surg 2008; 17:266-275.
2. Roberts DJ, Goldstein AM, Greame-Cook F, Dahms BB. Gastrointestinal tract and pancreas. In: Gilbert-Barness E, Kapur RP, Oligny LL, editors. Potter’s Pathology of the fetus, infant and child. 2nd ed. Philadelphia: Mosby; 2007. p. 1176-1178.
3. Martucciello G, Pini Prato A, Puri P, Holschneider AM, Meier-Ruge W, Jasonni V, et al. Controversies concerning diagnostic guidelines for anomalies of the enteric nervous system: a report from the fourth International Symposium on Hirschsprung's disease and related neurocristopathies. J Pediatr Surg  2005; 40:1527-1531.
4. Noviello C, Cobellis G, Romano M, Amici G, Martino A. Diagnosis of Hirschsprung's Disease: an age-related approach in children below or above one year. Colorectal Dis 2010; 12:1044-1048.
5. De Lorijn F, Kremer LC, Reitsma JB, Benninga MA. Diagnostic tests in Hirschsprung disease: a systematic review. J Pediatr Gastroenterol Nutr 2006; 42:496-505.
6. Rahman N, Chouhan J, Gould S, Joseph V, Grant H, Hitchcock R, et al. Rectal biopsy for Hirschsprung's disease--are we performing too many? Eur J Pediatr Surg  2010; 20:95-97.
7. Meier-Ruge WA, Bruder E. Current concepts of enzyme histochemistry in modern pathology. Pathobiology  2008; 75:233-243.
8. Nakao M, Suita S, Taguchi T, Hirose R, Shima Y. Fourteen-year experience of acetylcholinesterase staining for rectal mucosal biopsy in neonatal Hirschsprung's disease. J Pediatr Surg  2001; 36:1357-1363.
9. Kapur RP. Practical pathology and genetics of Hirschsprung's disease. Semin Pediatr Surg  2009; 18:212-223.
10. Kapur RP. Can we stop looking? Immunohistochemistry and the diagnosis of Hirschsprung disease. Am J Clin Pathol  2006; 126:9-12.
11. Baimbridge KG, Celio MR, Rogers JH. Calcium-binding proteins in the nervous system. Trends Neurosci 1992; 15:303-308.
12. McConalogue K, Low AM, Williamson S, Bornstein JC, Furness JB. Calretinin-immunoreactive neurons and their projections in the guinea-pig colon. Cell Tissue Res 1994; 276:359-365.
13. Barshack I, Fridman E, Goldberg I, Chowers Y, Kopolovic J. The loss of calretinin expression indicates aganglionosis in Hirschsprung's disease. J Clin Pathol  2004; 57:712-716.
14. Kapur RP, Reed RC, Finn LS, Patterson K, Johanson J, Rutledge JC. Calretinin immunohistochemistry versus acetylcholinesterase histochemistry in the evaluation of suction rectal biopsies for Hirschsprung Disease. Pediatr Dev Pathol  2009; 12:6-15.
15. Guinard-Samuel V, Bonnard A, De Lagausie P, Philippe-Chomette P, Alberti C, El Ghoneimi A, et al. Calretinin immunohistochemistry: a simple and efficient tool to diagnose Hirschsprung disease. Mod Pathol 2009; 22:1379-1384.
16. Pacheco MC, Bove KE. Variability of acetylcholinesterase hyperinnervation patterns in distal rectal suction biopsy specimens in Hirschsprung disease. Pediatr Dev Pathol  2008; 11:274-282.
Iranian Journal of Basic Medical Sciences
Volume 15, Issue 5 - Serial Number 5
September 2012
Pages 1053-1059

Files

Share

How to cite

Statistics