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Int J Clin Exp Pathol 2013;6(12):2955-2961

Original Article
Role of calretinin immunohistochemical stain in evaluation of Hirschsprung disease: an
institutional experience

Sanda Alexandrescu, Harvey Rosenberg, Nina Tatevian

Department of Pathology, University of California at San Francisco, CA, USA; Department of Pathology and Laboratory Medicine, University of
Texas at Houston, TX, USA

Received October 15, 2013; Accepted November 9, 2013; Epub November 15, 2013; Published December 1, 2013

Abstract: Background: The use of calretinin immunostain (IHC) in the evaluation of rectal suction biopsies for Hirschsprung disease (HD) has
been reported by Kapur et al. and others. The first goal of this article is to report our institutional experience with the use of calretinin in
specimens for evaluation of HD. The second goal is to describe the pattern of expression of calretinin in the junction of ganglionic-to-
aganglionic segment of pull through specimens of patients with a previous diagnosis of HD on suction rectal biopsy. Material and methods:
Three pathologists at University of Texas at Houston evaluated 28 rectal biopsy specimens from 2010-2011. The patients’ age ranged from 15
days to 8 years. Twenty-three cases were suction biopsies, and five were rectal full thickness biopsies. Hematoxylin-eosin (H&E) stain was
performed on at least 80 levels for the suction biopsy specimens. Calretinin immunohistochemical stain was performed on levels 40-42 in all
cases, with adequate controls. The H&E slides of nine pull through specimens with a diagnosis of HD on a suction rectal biopsy that was
evaluated in this study, were evaluated. Calretinin IHC was performed on the slide(s) showing the junction of aganglionic-to-normal rectum,
along with adequate controls. Results: The presence of ganglion cells consistently correlated with calretinin-positive thin nerve fibrils in the
lamina propria, muscularis mucosae and superficial submucosa. These nerve fibrils were absent in the aganglionic segments of bowel and in
the areas without ganglion cells from the junction of normal with diseased rectum. Calretinin was strongly expressed in the submucosal and
subserosal nerve trunks in the ganglionic segment. It had faint expression in the thick nerve trunks from the areas without ganglion cells 1.6-
2.5 cm proximal to the normal rectum. No calretinin expression was seen in the nerve trunks in the rest of the aganglionic segment.
Conclusion: The pattern of expression of calretinin in rectal suction biopsies in HD and normal rectum coincide with the ones previously
described in the literature. Calretinin IHC offered additional diagnostic value in the specimens with inadequate amount of submucosa and
rarely seen ganglion cells. The pattern of expression of calretinin in HD pull-through specimens correlates with the rectal biopsy ones. Faint
positivity of the thick submucosal and subserosal nerves in the absence of ganglion cells and calretinin positive nerve fibrils, is characteristic of
the junction of the aganglionic-to-normal rectum. We are the first ones to document this finding. (IJCEP1310037).

Keywords: Hirschsprung, calretinin, pull-through specimen

Address correspondence to: Dr. Sanda Alexandrescu, Department of Pathology, University of California at San Francisco, 505 Parnassus Ave,
M580, CA, USA. Tel: 713-992-7111; E-mail: sanda@erdani.com