Calretinin immunohistochemistry versus improvised rapid Acetylcholinesterase histochemistry in the evaluation of colorectal biopsies for Hirschsprung disease.
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Date
2014-07
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Abstract
Background: Acetylcholinesterase (AChE) histochemistry on rectal mucosal
biopsies accurately diagnoses Hirschsprung disease (HD), but is not widely
employed as it requires special tissue handling and pathologist expertise.
Calretinin immunohistochemistry (IHC) has been reported to be comparable
to AChE staining with the loss of expression correlating with aganglionosis.
Aim: The aim was to evaluate calretinin IHC as a primary diagnostic tool in
comparison to the improvised rapid AChE technique in the diagnosis of HD.
Materials and Methods: A total of 74 rectal biopsies (18 fresh frozen - 18 cases,
56 formalin fixed - 33 cases) from 51 cases of suspect HD were evaluated with
hematoxylin and eosin/AChE/Calretinin. Ten biopsies each from ganglionated
and aganglionated segments served as positive and negative controls. Ileal (3),
appendiceal (3) and ring bowel (2) biopsies were also included. Two pathologists
blinded to the clinical details evaluated the histomorphology with AChE and
calretinin. Observations were statistically analyzed and Cohen’s κ coefficient
employed to assess agreement between two pathologists and calretinin and the
AChE. Results: The study confirmed HD in 26 and non-HD in 25 cases. There
were 7 neonates, 5 low level biopsies and 14 “inadequate” biopsies. The results
of calretinin were comparable with AChE with a statistically significant measure
of agreement of κ = 0.973 between the two. One false-positive case of HD was
noted with calretinin. The advantages and disadvantages of calretinin versus
AChE are discussed. Conclusion: Calretinin is a reliable single immune marker
for ruling out HD by its specific positive mucosal staining of formalin fixed rectal
biopsy. The improvised AChE staining remains indispensable to confirm HD
on fresh biopsies and thus, along with calretinin IHC maximizes the diagnostic
accuracy of HD in difficult cases.
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Keywords
Acetylcholinesterase staining, calretinin, frozen sections, Hirschsprung disease, mucosal biopsies
Citation
Yadav Lokendra, Kini Usha, Das Kanishka, Mohanty Suravi, Puttegowda Divya. Calretinin immunohistochemistry versus improvised rapid Acetylcholinesterase histochemistry in the evaluation of colorectal biopsies for Hirschsprung disease. Indian Journal of Pathology & Microbiology. 2014 Jul-sept 57 (3): 369-375.