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An indirect competitive chemiluminescence enzyme immunoassay (icCLEIA) detection for the furaltadone metabolite 5-morpholinomethyl-3-amino-2-oxazolidone (AMOZ) was established. The effects of the coating antigen concentration, antibody dilution, immunoreaction time, reaction buffer and its ion concentration on the performance of the assay were studied and optimized. The results showed that the optimized assay conditions were as follows: the coating antigen concentration was 10 ng mL−1, the polyclonal antibody was diluted 60000 times, immunoreaction time was 50 min and the buffer solution was 0.01 M phosphate buffer solution (PBS, pH 7.4). Under the optimized conditions, the linear detection range of the developed icCLEIA was 0.026–3.52 μg L−1, the half inhibitory concentration (IC50) was 0.29 μg L−1 and the limit of detection (IC10) was 0.012 μg L−1. The average recoveries of AMOZ of spiked fish ranged from 101.0% to 115.5%. In conclusion, the icCLEIA is applicable for detection of trace AMOZ in fish samples. |
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