Calretinin Immunohistochemistry versus Acetylcholinesterase Histochemistry in the Evaluation of Suction Rectal Biopsies for Hirschsprung Disease

Kapur, Raj P.; Reed, Robyn C.; Finn, Laura S.; Patterson, Kathleen; Johanson, Judy; Rutledge, Joe C.

doi:10.2350/08-02-0424.1

Cited by 130 publications

(139 citation statements)

References 33 publications

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“…Of interest, recently Barshack et al 25 reported that calretinin immunostaining was absent in intrinsic nerve fibers in Hirschsprung colons, whereas it is identified in ganglion cells and intrinsic nerve fibers of nonHirschsprung patients. As a follow-up, Kapur et al 22 retrospectively applied calretinin immunostain to 14 Hirschsprung and 17 non-Hirschsprung patients, with staining results similar to the findings of Barshack et al 25 Further, this group compared the calretinin immunostain with the acetylcholinesterase stains performed at initial biopsy. They concluded that the calretinin immunohistochemical stain was superior to acetylcholinesterase histochemical stain in these evaluations.…”

Section: Discussionmentioning

confidence: 89%

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Utilization of peripherin and S-100 immunohistochemistry in the diagnosis of Hirschsprung disease

et al. 2010

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Evaluation of rectal biopsies for ganglion cells is performed for patients suspected of having Hirschsprung disease. At times, identification of ganglion cells can be difficult, especially in newborns. To assist in diagnosis, frozen tissue can be collected for acetylcholinesterase histochemical staining. At our institution, we developed a protocol using peripherin and S-100 immunostaining as an adjunct to hematoxylin and eosin (H&E) for the identification of ganglion cells. Further, at the time of frozen section, we performed Diff Quik staining to highlight ganglion cells. One hundred and thirty eight rectal biopsies submitted for evaluation of Hirschsprung disease were compiled from the archives of the Medical College of Georgia from 2002 to 2009. Initial evaluation consisted of eight levels of H&E-stained slides and two unstained slides each for immunostaining with peripherin and S-100. If on initial evaluation, ganglion cells were not identified, additional H&E and peripherin immunostains were performed. Peripherin immunostaining was unequivocally identified in the cytoplasm of ganglion cells of patients at all ages. Of the 136 patients with diagnostic biopsies, 80% had ganglion cells. Of these, 93% of cases were diagnosed on the original eight levels. Twenty-seven cases were devoid of ganglion cells, and of these, 81% showed submucosal neural hypertrophy on S-100 staining. Twenty-six patients had confirmed aganglionic segments at the time of colonic resection. One patient had colostomy only. A total of 54 frozen sections were performed on 25 patients over this same period of time. Diff Quick staining was found to be very useful. In this study, our protocol proved to be very sensitive, specific, and efficient for the diagnosis of Hirschsprung disease.

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Section: Discussionmentioning

confidence: 89%

“…In addition, the stains may be difficult to interpret, particularly in ambiguous cases. 3,4 The demands of interpretation of this stain are well documented with false positive, [4][5][6]8,9 false negative, [18][19][20][21][22] and equivocal 19,22 results.…”

Section: Discussionmentioning

confidence: 99%

Utilization of peripherin and S-100 immunohistochemistry in the diagnosis of Hirschsprung disease

et al. 2010

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“…21 Nerve cell bodies in both submucosa and myenteric ganglia of guinea pig 22,23 and in the human gastrointestinal tract are immunopositive for calretinin, a calcium-binding protein. 24 Recently, it has been suggested that calretinin might be more accurate than acetylcholinesterase in detecting aganglionosis 18 in a small series of rectal biopsy. Our study demonstrates for the first time in a large series of rectal suction biopsies that pathologists can diagnose all cases of HD with no false positives using the sole immunohistochemistry with calretinin antibody, and that the concordance between calretinin and the gold standard (ie, surgery for HD patients) is statistically excellent, whereas the concordance between the standard technique and the Gold standard is only good.…”

Section: Discussionmentioning

confidence: 99%

“…This binary pattern of interpretation (positive/negative) gives the technique distinctive ease and reproducibility, avoiding the need for repeated biopsies and delayed treatment. But because of the phenotypically heterogeneous nature of HD as provided by Kapur,18 the utility of calretinin immunohistochemistry, in particular in circumstances such as ultrashort segment HD or transitional zone assessments, remains to be determined, and calretinin immunohistochemistry should always be used in addition to limited hematoxylin and eosin sections. …”

Section: Discussionmentioning

confidence: 99%

Calretinin immunohistochemistry: a simple and efficient tool to diagnose Hirschsprung disease

et al. 2009

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Diagnosis of Hirschsprung disease (HD) is quite entirely based on the histopathological analysis of suction rectal biopsies. This hematoxylin and eosin approach has some limitations, despite the help of acetylcholinesterase staining. The aim of this study was to assess the diagnostic value of calretinin immunochemistry as a simple and reliable method in the diagnosis of HD. A total of 131 initial rectal biopsies carried out for suspicion of HD in children were retrieved, and calretinin immunohistochemistry was carried out on paraffin-embedded biopsies. Diagnosis of HD was made when no staining was observed. The results were statistically analyzed in comparison with our standard method (histology and acetylcholinesterase staining). 130 biopsies were accurately diagnosed on the basis of the positivity or negativity of calretinin staining. The senior pathologists diagnosed all cases of HD with no false positives. Furthermore, 12 additional cases initially considered as doubtful for HD using the standard method, were accurately diagnosed using calretinin immunohistochemistry. The false negative was a case of HD with a calretinin-positive biopsy. We also demonstrate the ease of calretinin interpretation compared with acetylcholinesterase for the junior pathologist. Calretinin immunohistochemistry overcomes most of the difficulties encountered using the combination of histology and acetylcholinesterase staining, and detects almost all cases of HD with confidence, with no false positives. Thus, we demonstrate that calretinin is superior to acetylcholinesterase to complete histology and could advantageously substitute for acetylcholinesterase. Modern Pathology ( The diagnosis of Hirschsprung disease (HD) is based on a combination of clinical features, radiological appearance of the bowel and histological features on suction rectal biopsies, 1,2 that is, aganglionosis and abnormally large nerves. In practice, a definite diagnosis of HD, leading to surgical treatment, is quite entirely based on histological results. 3,4 Several efficient approaches have been devised to assess the diagnosis of HD using a combination of standard histology and histochemistry. Standard histology obtained from rectal suction biopsies that sample mucosa and underlying submucosa, demonstrating rectal aganglionosis, requires the analysis of 100 or more histological sections to ensure good specificity, and is therefore time consuming. Moreover, difficulties in analysis may arise in several situations: (1) when the site of biopsy is too distal, because of the physiological paucity of ganglion cells; (2) when the sample is too superficial with not enough submucosa; and (3) when there is difficulty in identifying ganglion cells with confidence, particularly in neonates.For these reasons, standard histology is frequently supplemented with acetylcholinesterase histochemistry. 5,6 This technique, showing staining of extrinsic nerve fibers, provides quick results but requires frozen tissue samples. The last reports show good specificity of an abnormal ac...

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“…Acetylcholinesterase histochemistry (AChE) aids diagnosis but has its pitfalls as it requires special handling of the tissue. 7 One study further added that a certain type of nerve cell bodies in submucosa and the myenteric ganglia of the gastrointestinal tract were seen to show immunopositivity for calretinin. 8 Latestly, it is reported that calretinin immunohistochemical staining is found to be superior to conventional acetylcholinesterase staining to confirm absence of ganglions.…”

Section: Introductionmentioning

confidence: 99%

Calretinin Expression in Hirschsprung’s Disease – A Potential Marker of Ganglion Cells

Sikandar¹,

Nagi²,

Sikandar³

et al. 2017

APMC

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Calretinin Immunohistochemistry versus Acetylcholinesterase Histochemistry in the Evaluation of Suction Rectal Biopsies for Hirschsprung Disease

Cited by 130 publications

References 33 publications

Utilization of peripherin and S-100 immunohistochemistry in the diagnosis of Hirschsprung disease

Utilization of peripherin and S-100 immunohistochemistry in the diagnosis of Hirschsprung disease

Calretinin immunohistochemistry: a simple and efficient tool to diagnose Hirschsprung disease

Calretinin Expression in Hirschsprung’s Disease – A Potential Marker of Ganglion Cells

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