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.

Comparative study between use of calretinin and synaptophysin immunostaining in diagnosis of Hirschsprung disease

“…6 Hirschsprung disease (HD) is classically diagnosed or excluded by rectal biopsy, a method that take a sample a 2 to 3 mm of rectal mucosa and underlying submucosa. 7 Different approaches are customized to evaluate rectal biopsies, but the foundation for diagnosis or exclusion of HD in most practices is microscopic examination of hematoxylin and eosin (H&E)-stained sections. 8 High accurate identification of submucosal ganglion cells excludes HD, whereas aganglionic adequate sample is considered diagnostic.…”

“…This technique is applied in most pediatric pathology laboratories. 7 The reliability of this technique relys on the observer's ability to accurately identify a ganglion cell in H&E stained sections. Although no universal agreement regarding the number of histological sections needed to diagnose HD has been approved, previous works relied on the histopathological examination of 50 serial H&E stained sections.…”

Role of Calretinin in Determining the Aganglionic Segment in Hirschsprung's Disease

“…Tran et al (14) suggested that calretinin was more practical to determine the aganglionic segment in HSCR and that calretinin should be preferred first. In a study by Mukhopadhyay et al (15) with a larger patient group, calretinin was emphasized to be a very useful biomarker to distinguish HSCR from diseases with similar symptoms such as pseudo-HSCR, visceral neuropathy, and myopathy compared to immunomarkers such as cholinergic markers, neuropeptides, and synaptic markers.…”

Rectal Biopsy for Hirschsprung’s Disease: A Multicentre Study Involving Biopsy Technique, Pathology and Complications

“…Additional enzyme histochemistry staining with acetylcholinesterase (AChE) and NADH diaphorase has to be performed on fresh frozen specimens and is used in one hospital. Calretinin (positive in the control sample), S100 (positive in the control sample), synaptophysin (positive in the control sample) and AChE staining (negative in the control sample) highlight the nerve hypertrophy while NADH diaphorase and calretinin place the emphasis on the ganglion cells in the specimen [ 14 , 15 , 16 , 17 , 18 , 19 , 20 ].…”