Immunohistochemistry-based comparative study in detection of Hirschsprung’s disease in infants in a Tertiary Care Center

Abstract

BACKGROUND: Hirschsprung’s disease (HD) is the major cause of pediatric intestinal obstruction with a complex pattern of inheritance. The absence of ganglion cells along with an analysis of hypertrophy and hyperplasia of nerves in the nerve plexus of submucosa and muscularis mucosae is regarded as a potential hallmark for its diagnosis.

AIMS AND OBJECTIVES: This study was undertaken to ascertain the (1) clinical profile, (2) mode of presentation, and (3) to compare the role of calretinin immunostaining with acetylcholinesterase in the diagnosis of HD.

MATERIALS AND METHODS: This prospective and observational study was conducted in the Department of Pathology, IPGME & R from June 2014 to May 2015. One hundred and four patients clinically and radiologically diagnosed with HD underwent surgery were included in the study. The data of every patient including age, sex, and presenting symptoms were recorded. Eventually, histopathological, calretinin, and acetylcholinesterase immunohistochemical examination were done.

RESULTS: Total numbers of cases studied were 104, which aged between 0 days and 365 days. Male preponderance (76.92%) was noted. The overall sensitivity, specificity, positive, and negative predictive value of acetylcholinesterase were 100%, 86.44%, 84.91%, and 100%, respectively. The concordance of detection of ganglion cells and nerve fibers, and thereby diagnosis of Hirschsprung’s and non-HD using calretinin and the gold standard was statistically in strong agreement (κ = 0.749, 95% confidence interval: 0.635–0.863).

CONCLUSIONS: Calretinin stands out as the single and indispensable tool that differentiates HD from other mimickers.

Keywords

Acetylcholinesterase - calretinin - Hirschsprung’s disease - infants

Publication History

Received: 06 February 2016

Accepted: 29 June 2016

Article published online:
19 February 2020

• References

• 1 Dasgupta R, Langer JC. Hirschsprung disease. Curr Probl Surg 2004;41:942-88.
• 2 Newgreen D, Young HM. Enteric nervous system: Development and developmental disturbances – Part 2. Pediatr Dev Pathol 2002;5:329-49.
• 3 Newgreen D, Young HM. Enteric nervous system: Development and developmental disturbances – Part 1. Pediatr Dev Pathol 2002;5:224-47.
• 4 Suita S, Taguchi T, Ieiri S, Nakatsuji T. Hirschsprung's disease in Japan: Analysis of 3852 patients based on a nationwide survey in 30 years. J Pediatr Surg 2005;40:197-201.
• 5 Garver KL, Law JC, Garver B. Hirschsprung disease: A genetic study. Clin Genet 1985;28:503-8.
• 6 Badner JA, Sieber WK, Garver KL, Chakravarti A. A genetic study of Hirschsprung disease. Am J Hum Genet 1990;46:568-80.
• 7 Heitz PU, Komminoth P. Biopsy diagnosis of Hirschsprung's disease and related disorders. Curr Top Pathol 1990;81:257-75.
• 8 Lake BD, Risdon RA, Malone MT. Letter to the editor. Pediatr Surg Int 1988;3:373-4.
• 9 Babu MK, Kini U, Das K, Alladi A, D'Cruz AJ. A modified technique for the diagnosis of Hirschsprung disease from rectal biopsies. Natl Med J India 2003;16:245-8.
• 10 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;40:1527-31.
• 11 Athow AC, Filipe MI, Drake DP. Problems and advantages of acetylcholinesterase histochemistry of rectal suction biopsies in the diagnosis of Hirschsprung's disease. J Pediatr Surg 1990;25:520-6.
• 12 Moore SW, Johnson G. Acetylcholinesterase in Hirschsprung's disease. Pediatr Surg Int 2005;21:255-63.
• 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;57:712-6.
• 14 Meier-Ruge W. Histological diagnosis and differential diagnosis. In: Holschneider AM, Puri P, editors. Hirschsprung's Disease and Allied Disorders. Amsterdam: Harwood; 2000. p. 252-65.
• 15 De Lorijn F, Reitsma JB, Voskuijl WP, Aronson DC, Ten Kate FJ, Smets AM, et al. Diagnosis of Hirschsprung's disease: A prospective, comparative accuracy study of common tests. J Pediatr 2005;146:787-92.
• 16 Karim S, Hession C, Marconi S, Gang DL, Otis CN. The identification of ganglion cells in Hirschsprung disease by the immunohistochemical detection of ret oncoprotein. Am J Clin Pathol 2006;126:49-54.
• 17 Kini U, Babu MK, Yadav L, Mohanty S, Divya P, Saleem KM, et al. Role of syanptophysin in leveling circumferential full thickness (doughnut) bowel. Indian J Pathol Microbiol 2012;55:s10.
• 18 Kakita Y, Oshiro K, O'Briain DS, Puri P. Selective demonstration of mural nerves in ganglionic and aganglionic colon by immunohistochemistry for glucose transporter-1: Prominent extrinsic nerve pattern staining in Hirschsprung disease. Arch Pathol Lab Med 2000;124:1314-9.
• 19 Chentanez V, Chittmittrapap S, Cheepsoonthorn P, Agthong S. New classification of histochemical staining patterns of acetylcholinesterase activity in rectal suction biopsy in Hirschsprung's disease. J Med Assoc Thai 2000;83:1196-201.
• 20 Schemann M, Sann H, Schaaf C, Mäder M. Identification of cholinergic neurons in enteric nervous system by antibodies against choline acetyltransferase. Am J Physiol 1993;265(5 Pt 1):G1005-9.
• 21 Yadav L, Kini U, Das K, Mohanty S, Puttegowda D. Calretinin immunohistochemistry versus improvised rapid acetylcholinesterase histochemistry in the evaluation of colorectal biopsies for Hirschsprung disease. Indian J Pathol Microbiol 2014;57:369-75.