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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
Histochemical staining of rectal suction biopsies as the first investigation in patients with chronic constipation
Sandra Montedonico1, Anna Piaseczna Piotrowska, Udo Rolle
1The Children's Research Centre, Our Lady's Children's Hospital for Sick Children, Crumlin, Dublin 12, Ireland. sandramontedonico@hotmail.com
Insights
Rectal suction biopsy (RSB) is a safe and effective initial diagnostic tool for chronic constipation in children. Histochemical staining aids in accurately diagnosing conditions like Hirschsprung
Area of Science:
- Pediatric Gastroenterology
- Gastrointestinal Motility Disorders
- Histopathology
Background:
- Rectal suction biopsy (RSB) is the gold standard for diagnosing bowel motility disorders.
- Chronic constipation is a common pediatric issue requiring accurate diagnosis.
- Histochemistry enhances diagnostic capabilities of RSB.
Purpose of the Study:
- To evaluate the efficacy of RSB with histochemistry as a primary diagnostic method for pediatric chronic constipation.
- To assess the diagnostic accuracy for various intestinal dysganglionosis conditions.
Main Methods:
- Retrospective analysis of 766 children undergoing RSB for chronic constipation (1993-2005).
- Specimens stained with Hematoxylin and Eosin (H&E), Nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d), and Acetylcholinesterase (AChE).
- Analysis focused on submucosal tissue, ganglion cells, and nerve fiber staining patterns.
Main Results:
- Adequate submucosa obtained in 85.5% of cases.
- Hirschsprung's disease diagnosed in 7.2% (absent ganglion cells, increased AChE, lack of NADPH-d).
- Intestinal neuronal dysplasia identified in 9% (hyperganglionosis, giant/ectopic ganglia, increased AChE). Hypoganglionosis suspected in 1.3%.
Conclusions:
- RSB with histochemistry is a simple, safe, and accurate first-line diagnostic approach for pediatric chronic constipation.
- Combined NADPH-d and AChE staining significantly improves the diagnosis of intestinal dysganglionosis.
- RSB demonstrates high diagnostic yield for motility disorders in children.
Abstract:
Rectal suction biopsy (RSB) is the gold standard diagnostic procedure for disorders of bowel motility. This study describes our experience with RSB stained with histochemistry as the first diagnostic approach in a large series of patients presenting with chronic constipation. Between 1993 and 2005, 766 children underwent RSB for persistent chronic constipation. The specimens were snap frozen, sectioned and stained with conventional hematoxylin and eosin (H&E) and with nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) and acetylcholinesterase (AChE) histochemical stainings. Adequate amount of submucosa was present in 655 (85.5%) out of 766 cases and formed the basis of this study. RSB in 540 (82%) patients were reported as normal. Hirschsprung's disease was found in 47 (7.2%) patients with characteristic features of absence of ganglion cells, increased AChE activity in the lamina propria and muscularis mucosae, thick nerve fibers in the submucosa, and a lack of NADPH-d-positive fibers in muscularis mucosae. RSB in 59 (9%) patients presented features of intestinal neuronal dysplasia such as submucosal hyperganglionosis, giant ganglia, ectopic ganglia and increased AChE activity in lamina propria. Hypoganglionosis was suspected in nine (1.3%) children because of sparse or absent ganglion cells and low AChE and NAPDH-d activity in muscularis mucosae. Three patients (0.4%) developed bleeding following RSB, requiring diathermy of the bleeding point. Thus, we conclude that RSB is a simple and safe method when used as the first diagnostic approach in patients with chronic constipation. The combination of two histochemical stainings techniques provides a high level of accuracy in the diagnosis of intestinal dysganglionosis.
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