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Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein Gene Product 9.5
Published on: April 26, 2019
Pathogenesis of chronic constipation in a Polish group of paediatric patients - an attempt to create the optimal
Sylwia Szymańska1, Malgorzata Markiewicz-Kijewska2, Michal Pyzlak3
1Department of Pathology, The Children's Memorial Health Institute, Warsaw, Poland.
Insights
Diagnosing chronic constipation in children is challenging. Immunohistochemistry, particularly calretinin staining, aids in identifying ganglion cells but cannot replace expert pathological assessment for accurate diagnosis.
Area of Science:
- Pediatric Pathology
- Gastroenterology
- Histopathology
Background:
- Histopathological diagnosis of chronic constipation in children is challenging due to heterogeneous causes.
- Accurate diagnosis is crucial for effective treatment and management.
Purpose of the Study:
- To establish an optimal immunohistochemical (IHC) and histological diagnostic protocol for pediatric chronic constipation.
- To evaluate novel antibodies and analyze staining patterns for improved diagnostic accuracy.
Main Methods:
- Retrospective analysis of full-thickness intestinal biopsies from 28 pediatric patients.
- Inclusion of patients with Hirschsprung's disease (HD), desmosis of the colon (DC), chronic constipation of unknown etiology, and controls.
- Application of IHC stains (calretinin, CD117) and histological stains (picrosirius, trichrome gomori), analyzing staining patterns and intensity.
Main Results:
- Calretinin staining was positive in all patients with ganglion cells, aiding in their identification.
- No significant differences in staining intensity or pattern were observed between groups, except in HD where ganglion cells were absent.
- Calretinin staining did not unequivocally identify all ganglion cells compared to routine hematoxylin and eosin staining.
Conclusions:
- Calretinin is a valuable tool for identifying ganglion cells in pediatric intestinal biopsies.
- While helpful, calretinin staining does not replace the expertise of a pediatric pathologist.
- Full-thickness intestinal biopsies with additional IHC and histological stains, starting with picrosirius red, are recommended for diagnosing pediatric chronic constipation.
Introduction:
Histopathological diagnosis of chronic constipation in children is difficult and time-consuming because the aetiology of the problem is heterogenous.
Aim:
To create the optimal immunohistochemical (IHC) and histological diagnostic protocol using novel antibodies and assessing precisely their patterns.
Material And Methods:
Twenty-eight paediatric patients were enrolled to the study. The study group consisted of the following: 9 patients with confirmed Hirschsprung's disease (HD), 11 patients with desmosis of the colon (DC) (3) or with chronic constipation of unknown aetiology (3), and eight children operated on due to other problems. Retrospective analysis of full-thickness material from the large intestine was performed. In each specimen the number of ganglion cells was estimated per square millimetre as well as the number of submucosal and intramuscular ganglion cells per ganglion. The following IHC and histological stains were also performed: calretinin, CD117, picrosirius, and trichrome gomori. Patterns (nuclear vs. cytoplasmic vs. membranous) and intensity (strong vs. faint) of the stainings were analysed.
Results:
There was no statistically significant difference between groups while comparing the intensity and pattern of each staining, except HD (no staining due to lack of ganglion cells), p > 0.001. Calretinin was positive in each patient with ganglion cells; however, it did not unequivocally stain all cells identified in routine haematoxylin and eosin staining.
Conclusions:
Calretinin is helpful in identifying ganglion cells; however, it cannot replace an experienced paediatric pathologist. In children with chronic constipation it is worth obtaining a full thickness intestinal biopsy in order to perform additional histological and immunohistochemical stains starting with picrosirius red.
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