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Updated: Aug 23, 2026
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
Histological studies on Hirschsprung's disease and its allied disorders in childhood
Ryouichi Tomita1, Keimei Munakata, Edward R Howard
1First Department of Surgery, Nihon University School of Medicine, Japan. rtomita-ndus@mvg.biglobe.ne.jp
Insights
Accurate diagnosis of childhood constipation disorders like Hirschsprung
Area of Science:
- Pediatric Gastroenterology
- Histopathology
- Neurogastroenterology
Background:
- Chronic constipation, abdominal distension, and pain in children can indicate serious underlying conditions.
- Accurate histological diagnosis is crucial for managing Hirschsprung's disease (HD) and allied disorders like hypoganglionosis (Hypo) and intestinal neuronal dysplasia (IND).
- Distinguishing between these conditions is essential for appropriate treatment and patient outcomes.
Purpose of the Study:
- To evaluate the efficacy of rectal biopsy histology in diagnosing Hirschsprung's disease (HD) and its allied disorders in pediatric patients.
- To compare the diagnostic utility of acetylcholinesterase (AChE) and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining.
- To establish reliable histological methods for accurate diagnosis in children with refractory constipation.
Main Methods:
- Rectal biopsies were performed on 109 pediatric patients (aged 2-15 years) presenting with chronic refractory constipation.
- Tissue samples were stained using acetylcholinesterase (AChE) and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase.
- Histological examination focused on identifying ganglion cells and nerve fibers in mucosal and full-thickness specimens.
Main Results:
- Histological analysis diagnosed 20 cases of hypoganglionosis (Hypo), 5 of Hirschsprung's disease (HD), and 2 of intestinal neuronal dysplasia (IND).
- AChE staining accurately diagnosed HD and IND using rectal mucosal biopsies.
- NADPH-diaphorase staining of full-thickness rectal specimens was essential for the accurate diagnosis of Hypo.
Conclusions:
- Rectal mucosal biopsy with AChE staining is effective for diagnosing childhood HD and IND.
- NADPH-diaphorase staining in full-thickness rectal specimens provides accurate histological diagnosis for Hypo.
- NADPH-diaphorase staining facilitates easier detection of cholinergic fibers and ganglion cells compared to AChE staining.
Background/Aims:
To obtain accurate diagnosis for Hirschsprung's disease (HD) and its allied disorders such as hypoganglionosis (Hypo) and intestinal neuronal dysplasia (IND) in childhood patients with chronic constipation, we studied the histology of childhood patients with refractory constipation accompanied by abdominal distension and pain.
Methodology:
Based on clinical signs and symptoms noted on admission, all of 109 patients (60 males and 49 females, aged 2-15 years with a mean age of 9.8 years) were suspected to have chronic refractory constipation. To obtain accurate histological diagnosis in childhood patients with chronic refractory constipation, we performed rectal biopsies on these patients. Tissue samples were frozen and 12-microm sections were stained with acetylcholinesterase (AChE) by the method of Karnovsky and Roots, and with nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase by the modified Scherer-Singler's method.
Results:
On the basis of histological studies using rectal biopsies, 20 cases were diagnosed with Hypo, 5 with HD, 2 with intestinal neuronal dysplasia (IND), and 82 with normal findings. The incidence of normal cases was significantly greater than that of allied disorders of HD including both Hypo and IND (P<0.0001). The incidence of Hypo was also significantly greater than that of Hypo and IND (P<0.01, P<0.0001, respectively). Both HD and IND could be diagnosed by rectal mucosal biopsies with AChE staining. However, accurate diagnosis of Hypo could be made only through examination of the submucosal and myenteric plexuses by NADPH-diaphorase staining in full-thickness rectal specimens.
Conclusions:
We were able to obtain accurate diagnosis of childhood patients with HD and IND by rectal mucosal biopsy with AChE staining. On the other hand, accurate histological diagnosis in patients with Hypo could also be obtained by NADPH-diaphorase staining in full-thickness rectal specimens. That is to say, it is easier for the investigator to detect the cholinergic fiber and ganglion cell in the gut wall using NADPH-diaphorase staining than by using AChE staining.
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