Experience with a cholinesterase histochemical technique for rectal suction biopsies in the diagnosis of Hirschsprung's disease

Abstract

Cryostat sections from 160 rectal suction biopsies were stained for cholinesterases by the method of Karnovsky and Roots (1964) in an attempt to facilitate the diagnosis of Hirschsprung's disease.The method proved at least as reliable as experienced assessment of paraffin haematoxylin-eosin sections, and appeared to offer the advantages of reduced scanning fatigue and superior demonstration of the increased cholinesterase-positive nerves in Hirschprung's disease.Contrary to the findings of Meier-Ruge (1971) it was not possible to base a diagnosis on mucosal cholinesterase activity.

Histochemical diagnosis of Hirschsprung's disease and a comparison of the histochemical and biochemical activity of acetylcholinesterase in rectal mucosal biopsies.

“…The biopsies were trimmed to give a final specimen weighing less than 10 mg. The tissue was hand-homogenised in 0-1 M-phosphate buffer at pH 8-0 in a glass Potter-Elvehjem type homogeniser using 15 strokes. The volume of buffer was sufficient to give a final concentration of 10 mg tissue per ml.…”

“…Received for publication 8 October 1979 Table 1 Age and sex incidence ofpatients biopsied: numbers of cases of Hirschsprung's disease in parentheses Age group Males Females 0-4 weeks 7 (4) 0 (0) 1-12 months I I (0) 7 (1) 1-4 years 30 (4) 18 (1) 5-12 years 52 (6) 18 (0) 13-20 years 3 (1) 1 (0) > 21 years 1 (0) 2 (0) Totals 104 (15) 46 (2) The biopsies were taken by means of laryngeal punch biopsy forceps at several levels from the anal margin ( Table 2). The patients attended hospital as day cases on prearranged days of the week, and the procedure was carried out with sedation or under mild general anaesthesia.…”

Calretinin and microtubule-associated protein-2 (MAP-2) immunohistochemistry in the diagnosis of Hirschsprung's disease

“…We routinely performed 3 leveling sections and 10 step serial sections per specimen. Among them, 13 of 65 were inadequate specimens because they comprised only a mucosal layer without submucosal layers [11,12]. Therefore, we analyzed the 52 rectal suction biopsy specimens (37 from HD and 15 from non-HD) for the presence of ganglion cells with calretinin and MAP-2 immunohistochemistry (Table 1).…”

Hirschsprung’s Disease and Mimicking Conditions

“…Acetylcholinesterase activity in the lamina propria and muscularis mucosae can be shown with special staining [40], This is a dif ficult technique to master and an easier, more useful method is to estimate acetylcholines terase activity in a centrifuged specimen [41], A high level of activity will indicate Hirsch sprung's disease. Since false positives are pos sible [42], this should not be the only means of establishing the diagnosis.…”