Clinical Chemistry, Vol 29, 1065-1069, Copyright © 1983 by American Association for Clinical Chemistry

Rapid acetylcholinesterase screening test for neural tube defect

DL Hay, GF Ibrahim and I Horacek

A 5-min optimized automated assay for measuring acetylcholinesterase (AChE) and total cholinesterase (ChE) in amniotic fluids is described in which a centrifugal analyzer is used. Total ChE was measured by using acetylthiocholine iodide and monitoring the color developed after 5,5'-dithiobis(2-nitrobenzoic acid) is added. We evaluated four direct and indirect procedures for measuring AChE in which thiocholine is generated. These methods involve either the specific enzyme inhibitors ethopropazine and BW284C51 or the specific substrates S- butyrylthiocholine and acetyl-beta-methylthiocholine. We found the last method distinguishes normal pregnancies most efficiently from those with neural tube defect. Filtering samples through 0.22-micron membrane filters before analysis decreased the incidence of false positives. Significantly increased AChE concentrations were noted in all abnormal pregnancies (n = 45) and in 4.8% of normal pregnancies (n = 310). The rapid AChE assay involving acetyl-beta-methylthiocholine, when applied alongside a rapid alpha-fetoprotein assay, increases the efficiency of screening programs for pregnancies with neural tube defect to 99.4%. It also identifies samples with negative results for alpha-fetoprotein that require further examination by the sensitive gel-electrophoretic test for AChE.