Role of Calretinin Immunohistochemical Stain in Evaluation of Hirschsprung’s Disease

Abstract

Introduction: Hirschsprung’s Disease (HD) is a common congenital disorder characterised by absence of enteric neurons along a variable length of large bowel involving a segment of the rectum and the adjacent proximal bowel. The definitive diagnosis is based on histopathological study of rectal seromuscular biopsies. Calretinin Immunohistochemistry (IHC) staining adds to the diagnostic yield as it allows better appreciation of the mature and immature ganglion cells including the segments with paucity of ganglion cells. Aim: To assess the usefulness of calretinin IHC in the diagnosis of HD as an add-on technique to routine Haematoxylin and Eosin staining. Materials and Methods: This was a prospective study carried out in the Department of Pathology, SCB Medical College, Cuttack during the period from August 2016 to July 2018. Full thickness rectal biopsy specimens were processed and subjected to routine Hematoxylin and Eosin Stain (H&E) staining and Calretinin IHC staining. A thorough search of ganglion cells was made in both types of stained slides. The observations were tabulated and compared for diagnostic accuracy. Statistical analysis was done using Statistical Packages for Social Sciences (SPSS) Inc, Chicago IL version 15.0 windows. Through, ANOVA and Chi-square test. Results: Calretinin IHC provided superior diagnostic yield compared to routine H&E staining. A total number of 45 cases were studied. Sensitivity of 95.4% and specificity of 55.5% were obtained. It also presented higher accuracy values, than H&E (p=0.49). Conclusion: Calretinin is a superior immunohistochemical staining method that serves as a reliable additional diagnostic tool for better morphological appreciation of ganglion cells and thereby helps in establishing the diagnosis of HD.

Hirschsprung’s Disease Diagnosis: A Comparison of Calretinin and CD56 Immunohistochemistry

“…Histopathological analysis using standard H&E staining demonstrates that the Meissner's and myenteric plexuses are devoid of ganglion cells and that the submucosa of the aganglionic segment contains hypertrophied nerve fibers. 9 The intensity of ganglion cells declines at the end of the rectum, making it difficult to locate the ganglion cells. To overcome possible sampling issues, taking the biopsy at least 2 cm on top of the dentate line is advisable.…”

“…8 The presence of hypertrophied nerve fibers in the submucosa of the aganglionic segment and the absence of ganglion cells in the submucosal and myenteric plexuses are considered the gold standard for the diagnosis in histopathological examination of rectal biopsy utilizing routine standard hematoxylin and eosin (H&E) staining. 9 Several supplementary approaches, such as acetylcholinesterase (AChE) histochemistry and different immunohistochemistry (IHC) markers, have been developed to help with the diagnosis of HD because locating the ganglion cells can require several serials cut sections and have many drawbacks. 10 The AChE histochemistry approach has limitations, such as the need to treat frozen sections, interpretive issues, ambiguous or false-positive results, and technological obstacles.…”