Calretinin immunohistochemistry: A new cost-effective and easy method for diagnosis of Hirschsprung′s disease

Abstract

Aim:To evaluate the efficacy of calretinin immunostaining in diagnosing Hirschsprung's disease (HD).Materials and Methods:Sixty cases were studied over a period of 1 year (July 2010-June 2011). There were 36 full-thickness biopsies and 24 resected specimens. Calretinin processing was done on the paraffin-embedded blocks after routine histopathological examination.Results:Of the 36 biopsy specimens, in 19 cases HD was diagnosed by hematoxylin and eosin (H and E) staining earlier. In 2 patients, ganglion cells were seen and HD was ruled out. In 15 cases, there was a diagnostic dilemma and calretinin was used. Ganglion cells were found in 3 specimens and nerve fibers in 5. In all 24 resected specimens, calretinin correlated with the findings on H and E staining.Conclusions:Calretinin was extremely useful in solving the suspicious and indeterminate cases of HD. It can serve as a valuable cost-effective diagnostic aid in the centers where acetylcholinesterase enzyme histochemistry is not available.

The zebrafish mutant lessen: an experimental model for congenital enteric neuropathies

“…In HSCR patients, no CRT expression was observed in the aganglionic intestine, whereas CRT was detected in both ganglion cells and nerve fibers in the normoganglionic regions, leading to the conclusion that the absence of CRT in intestinal biopsies may be used as a diagnostic tool. [50][51][52][53] We further observed in lsn mutants that the expression of VIP and PACAP, which in zebrafish completely colocalize in enteric nerve fibers, 10,14 was disturbed. Previous work has revealed that nitrergic neurons of the zebrafish intestine coexpress VIP and PACAP.…”

“…Lsn embryos exhibited a decreased number and proportion of CBP‐expressing neurons in comparison with wt animals. In HSCR patients, no CRT expression was observed in the aganglionic intestine, whereas CRT was detected in both ganglion cells and nerve fibers in the normoganglionic regions, leading to the conclusion that the absence of CRT in intestinal biopsies may be used as a diagnostic tool …”

Calretinin-Immunoreactive Mucosal Innervation in Very Short-Segment Hirschsprung Disease: A Potentially Misleading Observation

“…It has been suggested [16] that calretinin immunostaining alone may be sufficient to exclude HD in biopsies with inadequate submucosa. However, descriptions of infrequent patients with HD and intact CRir mucosal innervation in suction biopsies (false-negative diagnostic result) [8,11,12,15] raise concern that CRir distal projections from submucosal ganglion cells may extend into aganglionic bowel and complicate diagnosis of HD, particularly in patients with vsHD.…”

“…Calretinin is a calcium binding protein found in a subset of enteric neurons, including the processes of submucosal ganglion cells, which innervate the mucosa [9]. Several studies [8,[10][11][12][13][14][15][16] have shown that calretininimmunoreactive (CRir) mucosal innervation is absent in the aganglionic segment of HD and that this finding is as *Corresponding author, e-mail: raj.kapur@seattlechildrens.org Pediatric and Developmental Pathology 17, 28-35, 2014 DOI: 10.2350/13-10-1387-OA.1 ª 2014 Society for Pediatric Pathology diagnostically reliable as, if not superior to, AChE histochemistry. Although the triad of aganglionosis, submucosal nerve hypertrophy, and absent CRir mucosal innervation in a suction rectal biopsy may be diagnostic of HD, deviation from this pattern can lead to uncertainty.…”

Evaluation of calretinin immunohistochemistry as an additional tool in confirming the diagnosis of Hirschsprung disease

“…The stain has only recently gained broader recognition and has become the most common adjunct immunohistochemical marker used to facilitate the diagnosis of HD. The stain is easy to interpret and works with paraffin embedded material [13][14][15][16]. Numerous studies comparing the results of calretinin immunohistochemistry with those of AChE histochemistry demonstrate that calretinin may be superior to AChE assay [1,[17][18][19].…”