title: Development of an indirect competitive ELISA for simultaneous detection of enrofloxacin and ciprofloxacin.
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 (IC(50)) 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.
authors: College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China.
authors: Hai-tang
authors: HT
authors: Zhang
authors: Jin-qing
authors: JQ
authors: Jiang
authors: Zi-liang
authors: ZL
authors: Wang
authors: Xin-yao
authors: XY
authors: Chang
authors: Xing-you
authors: XY
authors: Liu
authors: San-hu
authors: SH
authors: Wang
authors: Kun
authors: K
authors: Zhao
authors: Jin-shan
authors: JS
authors: Chen
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000032:analysis*; D002417:Cattle; D002939:Ciprofloxacin / Q000032:analysis*; D000077422:Enrofloxacin; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D024841:Fluoroquinolones / Q000032:analysis*; D057230:Limit of Detection; D008460:Meat / Q000032:analysis*; D011817:Rabbits; D013552:Swine
chemical_list: D000890:Anti-Infective Agents; D024841:Fluoroquinolones; D000077422:Enrofloxacin; D002939:Ciprofloxacin