Indirect Competitive Chemiluminescence Enzyme Immunoassay for Furaltadone Metabolite in Metapenaeus Ensis
Abstract An indirect competitive chemiluminescence enzyme immonoassay (icCLEIA) for the quantitative analysis of furaltadone metabolite 5-morpholinomethyl-3-amino-2-oxazolidone(AMOZ) in Metapenaeus ensis sample was established with the highly sensitive luminol-H 2 O 2 -HRP-4-iodophenol chemiluminescence reaction detection system. The icCLEIA method was based on a single-chain variable fragment (scFv) antibody against AMOZ derivative. The optimized assay conditions were as follows: 62.5 μg L −1 of coating antigen (5-morpholinomethyl-3-(glyoxalamino)-2-oxazolidone-ovalbumin, AMOZA-OVA) was used in the experiment, the dilute multiple of scFv antibody was 1:10, the immunoreaction time was 45 min, and the dilute multiple and incubation time of HRP-goat anti mouse IgG were 1:10000 and 50 min, respectively. The icCLEIA results showed that IC 50 value and limit of detection (LOD) were 1.38 μg L −1 and 0.09 μg L −1 , and linear range was in the range of 0.26–9.1 μg L −1 . Inter-assay and intra-assay RSD were both below 15%. The scFv antibody showed high specificity. The average recoveries of four spiked level of AMOZ were 72.2%, 73.4%, 72.6% and 78.6%, respectively. In comparison with HPLC-MS/MS, there was a good correlation between the two methods ( R 2 = 0.9997). The established icCLEIA method could be further used for detecting AMOZ in aquatic products samples rapidly and efficiently.