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| Development of an indirect competitive ELISA for simultaneous detection of enrofloxacin and ciprofloxacin |
| Hai-tang Zhang, Jin-qing Jiang, Zi-liang Wang, Xin-yao Chang, Xing-you Liu, San-hu Wang, Kun Zhao, Jin-shan Chen |
| College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China, Henan Higher Education Engineering Technology Research Center for Animal Diseases Control and Residues Supervision, Xinxiang 453003, China |
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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 (R2=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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Received: 12 March 2011
Published: 04 November 2011
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