ZHU Libin,ZHANG Hua,LIN Jinhan, et al. Diagnosis of Hirschsprung disease with modified NADPH-d and AchE enzyme histochemistry[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2014, 44(1): 31-34.
Abstract：Objective: To assess the superiorities of modified NADPH-diaphorase (NADPH-d) and acetylcholinesterase (AchE) enzyme histochemistry staining technique to diagnose Hirschsprung disease (HD). To increase the identification of ganglion cells in the boundary of the bowel in HD and decrease the risks of reoperation. Methods: Full-thickness colon samples of 18 children with HD (girl 7 cases, boy 11 cases ), with average age (8.6±3) months, were examined. Frozen sections at three levels (narrow segment, transitional segment and dilated segment) were performed and investigated to determine the extent of aganglionosis by modified NADPH-d and AchE staining respectively, which were compared by traditional HE staining. TUJ-1 (β-III Tubulin) and PGP9.5(Protein Gene Product 9.5), markers of enteric neural cells, were used for identification of positive myenteric plexus and ganglion cells by immunofluorescence or immunohistochemistry. Results: NADPH-d staining could rapidly confirm whether the normal ganglion cells existed at the proximal end of the resected bowel. AchE technique clearly stained submucosal hypertrophic nerve trucks in aganglion segments. The specifica identifications of HD, NADPH-d staining was 94.4% (17/18), and AchE staining was 88.9% (16/18), respectively (P>0.05). Time comsuming was (7.0±1.7) min in NADPH-d, and (30.9±2.2) min in AchE, respectively (P<0.01). The results of two modified methods mentioned above were in keeping with TUG-landPGP 9.5. Conclusion: Modified NADPH-d staining can rapidly confirmed whether the ganglion cells distribute at proximal end of resected bowel and it is suitable for rapidly intraoperative diagnosis of HD. AchE staining is the best technique to demonstrate hypertrophic nerve trunks in the submucosa and is suitable for diagnosing samples of suction rectal biopsy.
