A sensitive, rapid chemiluminescence ELISA for the detection of 1, 4-bisdesoxycyadox residue in chicken muscle and liver

Abstract

A rapid system for 1, 4-bisdesoxycyadox (Cy4) detection in chicken breast muscle and liver using a highly sensitive luminol hydrogen peroxide-based chemiluminescence enzyme-linked immunosorbent assay (CL-ELISA) was developed. Cy4-specific monoclonal antibodies (Mab) were prepared. The Cy4 Mab dilution ratios, coating antigen (Cy4–OVA) concentration, blocking concentration, immunoreaction time and CL reaction time were optimized. The specificity of the assay was assessed by determining the cross-reactivity with the Cy4 parent drug Cyadox (CYX) and its related analogues and metabolites. For this method, the limit of detection was 0.007 ng mL⁻¹, the linear range (IC₂₀–IC₈₀) was 0.027–67.673 ng mL⁻¹, the intra-assay and inter-assay precisions were <10% and <15%, respectively, and the recovery was 65.46–101.33%. When chicken breast muscle and liver were examined by CL-ELISA and high performance liquid chromatography, the results of the two techniques correlated well, indicating that the CL-ELISA method was suitable for the screening and quality detection of veterinary drug residues in animal tissues.