Abstract
BackgroundHematoxylin-eosin (HE) staining of a full-thickness rectal wall fragment is classically used for the diagnosis of Hirschsprung disease (HD). However, this technique requires large fragments for a better diagnosis. Additionally, the histochemical and immunohistochemical methods of staining small fragments of rectal mucosal and submucosal biopsies are not available in all centers. Therefore, the possibility of diagnosing HD through HE staining in these biopsies could be a valuable alternative for centers that do not have more specific techniques. The objectives of the current investigation were to evaluate the concordance of the results obtained by HE staining and the calretinin method with acetylcholinesterase (AChE) activity in fragments of mucosa and submucosa in the diagnosis of HD.MethodsFor this study, 50 cases from our laboratory were selected. The tissue material was embedded in paraffin. Sixty levels of each fragment were utilized for HE, and the other 3 levels were used for calretinin. These slides were analyzed under the microscope, photographed and classified as either positive for HD when no ganglion cells were found with nerve trunks present or as negative when ganglion cells were found. The results from reading the slides were compared with those of AChE.ResultsOf the 50 cases evaluated by the HE technique, only 5 contradicted the diagnosis based on AChE, with a Kappa value of 0.800 and an accuracy of 90%. In the comparison between calretinin and AChE, 8 cases were discordant, with a Kappa value of 0.676 and an accuracy of 84%.ConclusionsThe concordance of results from AChE and HE methods was satisfactory, allowing for the potential use of the HE method for fragments of mucosa and submucosa as a valid alternative in the diagnosis of HD. The immunohistochemical technique of calretinin did not show good agreement with the AChE activity in our study.
Highlights
Hematoxylin-eosin (HE) staining of a full-thickness rectal wall fragment is classically used for the diagnosis of Hirschsprung disease (HD)
Hirschsprung disease (HD) is a congenital disease that is characterized by the lack of enteric neuron formation during the embryonic period, with the absence of ganglion cells and hypertrophy of nerve trunks in the terminal segment of the large intestine [1,2,3]
We investigated the accuracy of the calretinin assay for the diagnosis of HD and compared both techniques to the AChE assay
Summary
Hematoxylin-eosin (HE) staining of a full-thickness rectal wall fragment is classically used for the diagnosis of Hirschsprung disease (HD). This technique requires large fragments for a better diagnosis. The histochemical and immunohistochemical methods of staining small fragments of rectal mucosal and submucosal biopsies are not available in all centers. Hirschsprung disease (HD) is a congenital disease that is characterized by the lack of enteric neuron formation during the embryonic period, with the absence of ganglion cells and hypertrophy of nerve trunks in the terminal segment of the large intestine [1,2,3]. A biopsy of the rectal wall is the method of choice for diagnosis because of the pathological characteristics of HD. The best histological method for staining the rectal biopsy samples remains controversial [4,5,6]
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