Abstract
Modified 1-ethyl-3-(3-dimethylaminopropy) carbodiimide (EDC) method was employed to synthesize the artificial antigen of enrofloxacin (ENR), and New Zealand rabbits were used to produce anti-ENR polyclonal antibody (pAb). Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay (ELISA) standard curve was established. This assay was sensitive and had a linear range from 0.6 to 148.0 μg/kg (R 2=0.9567), with the half maximal inhibitory concentration (IC50) and limit of detection (LOD) values of 9.4 μg/kg and 0.2 μg/kg, respectively. Of all the competitive analogues, the produced pAb exhibited a high cross-reactivity to ciprofloxacin (CIP) (87%), the main metabolite of ENR in tissues. After optimization, the matrix effects can be ignored using a 10-fold dilution in beef and 20-fold dilution in pork. The overall recoveries and coefficients of variation (CVs) were in the ranges of 86%–109% and 6.8%–13.1%, respectively. It can be concluded that the established ELISA method is suitable for simultaneous detection of ENR and CIP in animal tissues.
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Project supported by the Henan Innovation Project for University Prominent Research Talents (No. 2010HASTIT026), and the Key Scientific & Technological Project of Education Department in Henan Province of China (No. 2011A230003)
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Zhang, Ht., Jiang, Jq., Wang, Zl. et al. Development of an indirect competitive ELISA for simultaneous detection of enrofloxacin and ciprofloxacin. J. Zhejiang Univ. Sci. B 12, 884–891 (2011). https://doi.org/10.1631/jzus.B1100076
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DOI: https://doi.org/10.1631/jzus.B1100076