University of Khartoum

Assessment of Calretinin Immunohistochemistry As A Diagnostic Tool In Hirschprung’s Disease Among Sudanese Patients In Soba University Hospital

Assessment of Calretinin Immunohistochemistry As A Diagnostic Tool In Hirschprung’s Disease Among Sudanese Patients In Soba University Hospital

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Title: Assessment of Calretinin Immunohistochemistry As A Diagnostic Tool In Hirschprung’s Disease Among Sudanese Patients In Soba University Hospital
Author: Mohamed, Alaa Hatim Ameer
Abstract: Background and Objective: Hirschsprung’s disease (HSD) is a common congenital disorder that results from absence of parasympathetic ganglion cells in the intramural and submucosal plexuses. It is the most common cause of organic constipation worldwide and it diagnosed early in life. The diagnosis is mainly based on histopathological examination of suction biopsies. This study aims at evaluating calretinin expression, and its specificity and sensitivity in different bowel segments and biopsies as a cost effective and more accurate method for diagnosis of HSD compared to the conventional H&E stain. Calretinin is a calcium binding protein that shows strong nuclear and cytoplasmic staining of ganglion cells and nerve fibers and negative staining in HSD. Materials and Methods A sample of 100 patients was obtained from Soba University Hospital during January 2013- December 2017. Of these, 98 were diagnosed with HSD histopathologically with different types of biopsies (44 suction rectal biopsies, 42 pull through operations, (20 narrow segments, 22 dilated segments), and 14 colostomy site biopsy). Additional 3 samples near the dentate line were considered as control. H&E slides were reevaluated and calretinin immunostain was performed on paraffin embedded blocks and seen by the researcher, supervisor and co-supervisor.   Results: 100 cases were evaluated, their age range (1 day-23 yrs), the most common age group for HSD is 1 day to 5 years (72%), with male to female ratio of (4.8:1). The rectum is the most common site of involvement (44%) followed by rectosigmoid colon (28%). Chronic constipation turned out to be the most common clinical presentation (49%), followed by abdominal distension (13%) and delayed meconium passage (8%). In 61% cases diagnosis of HSD is based on clinical features, radiological studies and histopathological evaluation for absence of ganglion cells. Of 44 rectal suction biopsies examined, H&E showed presence of ganglion cells in 8 (18.2%) and absence in 36 (81.8%), while calretinin showed positive staining in 3 (6.8%), negative in 39 (88.6%) and weak positive staining in 2 (4.5%) which are considered immature ganglion cells, (p= 0.03). In the 42 pull through bowel segments, H&E showed presence of ganglion cells in 21 (50%) and absence in 21 (50%), while calretinin showed positive staining in 20 (47.6%) and negative staining in 22 (52.4%), (p= 0.00). Of the 14 colostomy biopsies evaluated, 12 (85.7%) showed presence of ganglion cells and 2 (14.3%) absence of ganglion cells, while calretinin showed positive staining in 13 (92.9%) and negative in 1 (7.1%), (p= 0.011). Nerve fibers were evaluated in all samples examined, 7showed weak positive expression for calretinin and were considered as transitional zone between ganglionic and aganglionic segments. Calretinin showed very high sensitivity and specificity compared to H&E stain in all samples examined. In suction biopsies, calretinin showed 95.1% sensitivity and 100% specificity while H&E showed 85.3% sensitivity and 66% specificity. In pull through and colostomy biopsies, calretinin showed 100% sensitivity and 100% specificity while H&E showed 93.9% sensitivity and 91.3% specificity. Conclusion: Calretinin immunohistochemistry is not only cost effective, but is also a more accurate method for diagnosis of HSD in all types of biopsies with very high sensitivity and specificity and no false positive or negative results.
Description: 141 Pages
URI: http://khartoumspace.uofk.edu/123456789/27129
Date: 2018


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