Development of an indirect competitive enzyme-linked immunosorbent assay for screening ethopabate residue in chicken muscle and liver?

RSC Advances Pub Date: 2017-07-20 DOI: 10.1039/C6RA20736G

Abstract

Ethopabate (ETP) is a coccidiostat that is frequently used to prevent and treat coccidiosis in chickens. Illegal use and abuse of ETP can lead to drug residues in edible animal tissues, which present a potential health risk to consumers. To rapidly monitor ETP residues, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) has been developed that has simple sample preparation and clean-up. After immunogen preparation, inoculation and cell fusion, a monoclonal antibody (4G7) was obtained with the lgG2 isotype. The 4G7 antibody had the ability to specifically recognize ETP with an IC50 value of 0.66 μg L?1, it was found to have weak and insignificant cross-reactivity for some structure-related analogs. With the optimized ic-ELISA protocol, the detection limits of ETP were calculated as 0.21 μg kg?1 and 0.34 μg kg?1 in chicken muscle and liver samples, respectively. The recoveries ranged from 85.4% to 98.4% with a coefficient of variation of less than 15%. Furthermore, good correlations between the results of ic-ELISA and high-performance liquid chromatography demonstrated the reliability of the developed ic-ELISA. This proposed method is a rapid, sensitive and useful tool that offers a cost-effective alternative to current published procedures without any concession in the ability to detect ETP residues in edible animal tissues.

Graphical abstract: Development of an indirect competitive enzyme-linked immunosorbent assay for screening ethopabate residue in chicken muscle and liver
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