Evaluation and quality control of a monoclonal antibody based enzyme antigen immunoassay of acetylcholinesterase in amniotic fluid.

Abstract

We have evaluated a new monoclonal antibody based enzyme antigen immunoassay (EAIA) for acetylcholinesterase in amniotic fluid, intended for the detection of open fetal abnormalities, especially open neural tube defects. With a sample volume of 50 microliters, the detection limit is 0.7 nkat/l and linearity is found up to an amount 200-times the detection limit. CV's are less than or equal to 6.6% within assays and less than or equal to 10.3% between assays; recoveries averaged 103%. The upper limit of the reference range for amniotic fluids from non-affected pregnancies is 8.5 nkat/l for clear specimens and 25.0 nkat/l for haemolysed specimens. Amniotic fluid specimens from 1473 patients (1388 normal and 85 pathological pregnancies) were examined with both the new EAIA and the original procedure using a polyclonal rabbit antiserum. The two procedures showed identical sensitivities and specificities of about 100%, except for amniotic fluid samples containing haemolysed blood. For the latter the new EAIA showed a specificity of 100% compared to 55.6% for the original procedure. We conclude that the new EAIA is accurate and reproducible and shows, compared with the original procedure, an increased specificity in the analysis of amniotic fluid samples containing haemolysed blood.