Comparison of Neuropeptide Y, Protein Gene Product 9.5, and Acetylcholinesterase in the Diagnosis of Hirschsprung's Disease

Abstract

We compared the results of acetylcholinesterase (AChE) staining of mucosal rectal biopsy specimens with those using neuropeptide Y (NPY) and protein gene product 9.5 (PGP9.5) in biopsies from 68 patients. Thirty-three did not have Hirschsprung's disease (HD), 28 had proven HD, and biopsies from 7 patients had shown a slight increase in AChE stain but the patients did not have HD. In our hands, AChE stain was superior to the other two; it was easier to read and gave the most accurate results with no false-positive cases and only two instances in which the findings were suggestive but not diagnostic. Neuropeptide Y and PGP9.5 have the advantage that they can be used in paraffin-embedded material. With NPY, the results are closer than with PGP9.5 to those obtained with the AChE. Protein gene product 9.5 had the highest incidence of false-positive and false-negative results, but it stains nerve fibers and all neurons intensely and may be useful in the assessment of increased or decreased amounts of neural elements in the bowel.

Diagnosis of Hirschsprung Disease Using the Rectal Biopsy: Multi-institutional Survey

“…As an adjunct to morphologic examination, the AChE assay remains a superior histochemical assay [17] for the confirmation of HD relative to various immunohistochemical markers [1,2,17]. The staff at Children's Hospital Medical Center, Cincinnati, have demonstrated that a combination of well-controlled AChE staining and morphologic review of a sufficient RB will provide a diagnostic yield of the presence or absence of HD in 99% of cases; cases insufficient for AChE examination contained only anorectal mucosa or absence of muscularis mucosae.…”

“…2) in which single positively stained submucosal nerves can extend into the muscularis mucosae associated with several intramuscular thread-like nerve fibers. A well-developed normal AChE pattern may be confused with a less pronounced abnormal AChE pattern [3,17], leading to a false-positive [3] or equivocal [17] HD diagnosis. Inflammation may also exaggerate the normal AChE pattern (see Table 3 and its discussion).…”

“…The practice patterns of these institutions, which have all contributed substantially to the HD medical literature [1][2][3][5][6][7][8], are analyzed in an effort to offer guidelines to the practicing pathologist in approaching HD diagnosis by rectal biopsy. It must be stressed that we offer only practice guidelines and not absolute criteria as to practice parameters, as these may vary even among the institutions cited in this paper-a point that will be emphasized throughout.…”

Hirschsprung Disease and Other Enteric Dysganglionoses

“…74 A variety of alternative immunohistochemical markers (e.g., S100, NPY, PGP9.5, microtubule-associated proteins) have been used to identify abnormal nerve fibers in HSCR, 55,56,69,72,[76][77][78] but in comparative studies AChE histochemistry was deemed superior to NPY, PGP9.5, or S100 immunolabeling. 55,79 The current protocol in many laboratories is to request at least two suction biopsies from a site 2 cm above the pectinate line. 51 One of the samples is frozen in embedding medium for AChE staining and the other is fixed and embedded in paraffin.…”

Mucosal nerve deficiency in chronic childhood constipation: a postmigration defect?

“…Furthermore, moderate to mild mucosal nerve deficiency tended to predominate in the proximal segments of HSCR transitional zones that contained ganglia. Galvis and Yunis [20] described sparse mucosal innervation in "non-Hirschsprung" rectal biopsy specimens in paraffin-embedded sections without further comment. Their main focus was comparison of PGP 9.5, acetylcholine esterase, and neuropeptide Y as the preferable method in conjunction with H&E staining for diagnosis of HSCR.…”