Estimation of plasma and RBC acetylcholinesterase in children: An evaluation tool for Hirschsprung's disease?

Background: Increased acetylcholinesterase (AChE) activity on frozen sections of rectal mucosal biopsies accurately diagnoses Hirschsprung disease (HD). But the quest for a biomarker in blood as a screening test prompts one to look for AChE in blood and study its role in HD diagnosis. Aim: To develo...

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Published in:	Indian journal of pathology & microbiology Vol. 64; no. 2; pp. 266 - 276
Main Authors:	Kuvelkar, Neha, DSouza, Serena, Vidhyashree, K, Shankar, Gowri, Bukel, Maria, Noronha, Santosh, Kini, Usha
Format:	Journal Article
Language:	English
Published:	India Wolters Kluwer India Pvt. Ltd 01-04-2021 Medknow Publications and Media Pvt. Ltd Medknow Publications & Media Pvt. Ltd Wolters Kluwer Medknow Publications
Subjects:	Acetylcholinesterase Acetylcholinesterase - analysis Acetylcholinesterase - blood Acetylcholinesterase - metabolism acetylcholinesterase enzyme assay Biomarkers Biomarkers - analysis Biomarkers - blood Biopsy Child Child, Preschool Children Double-Blind Method ellman's method Erythrocytes Erythrocytes - enzymology Ethylenediaminetetraacetic acid Gastrointestinal Transit - physiology High-definition television Hirschsprung Disease - blood Hirschsprung Disease - diagnosis Hirschsprung Disease - pathology Hirschsprung's disease Histochemistry Histocytochemistry - methods Humans India Medical screening Mucosa Mucous Membrane - enzymology Plasma Prospective Studies Rectum Rectum - enzymology red blood cells Sensitivity and Specificity India Hirschsprung's disease red blood cells Acetylcholinesterase enzyme assay plasma Ellman's method
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Summary:	Background: Increased acetylcholinesterase (AChE) activity on frozen sections of rectal mucosal biopsies accurately diagnoses Hirschsprung disease (HD). But the quest for a biomarker in blood as a screening test prompts one to look for AChE in blood and study its role in HD diagnosis. Aim: To develop a low-cost reliable method to estimate the AChE activity in plasma and red blood cells (RBCs) in normal children (control) and study its role in HD (test). Materials and Methods: Optimized method derived after modifying and standardizing known AChE assay protocols for blood were employed on 30 controls to define the AChE cut-off range, on 40 suspected HD cases to categorize them as HD/non-HD based on cut-off values and later compared with gold standard tissue AChE histochemistry of rectal mucosal biopsies. Results: An optimal in-house modified methods of Ellman's was found best suited to analyze plasma AChE activity, method by Wilson and Henderson was optimal for extraction and AChE estimation in RBCs. AChE levels (controls) obtained were 1.03 ± 0.31 U/mL and 5.17 ± 1.52 U/mL in plasma and RBCs, respectively while the plasma AChE was 1.35 ± 0.84 U/mL (HD) and 1.62 ± 0.85 U/mL (non-HD) while RBC AChE was 4.29 ± 3.2 U/mL (HD) and 6.48 ± 4.31 U/mL (non-HD). Sensitivity was 66.67% and 55.56%, specificity was 22.73% and 45.45%, and an accuracy rate of 42.5% and 50% for plasma and RBC, respectively. Conclusions: Mutually exclusive AChE activity range identified for test blood samples overlapped with the normal and hence, not considered a diagnostic tool for HD.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0377-4929 0974-5130
DOI:	10.4103/IJPM.IJPM_567_19