Development of an Indirect Competitive ELISA for Rapid Analysis of Furazolidone Metabolite Residues in Animal Food
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Abstract:
Based on artificial antigen and specific polyclonal antibody, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed to detect 3-amino-2-oxazolidone (AOZ), followed by derivatization with 2-nitrobenzaldehyde. The selection of the optimum concentrations of the coating antigen, the polyclonal antibody and HRP-conjugated antibody was performed by ic-ELISA and square titration. Calibration curve was prepared for NPAOZ (3-(2-Nitrobenzylidenamino)-2-oxazolidinone), a 2-nitrobenzaldehyde derivative of AOZ, and good linearity was achieved over the concentration (0.25-10 ng/mL), the obtained IC50 was between 0.5881 ng/mL and 1.1333 ng/mL. In addition, the antibody was high specific for NPAOZ and no cross-reactivity with analog. The accuracy of the assay was validated by determining the recovery in shrimp tissue. High temperature and short time derivative process can be in line with incubating at 37 ℃ overnight.
