Development of a highly sensitive and specific immunoassay for enrofloxacin based on heterologous coating haptens

• A new derivative of enrofloxacin was synthesized and used as coating hapten.
• The effect of coating hapten on the sensitivity and specificity of ELISA was studied.
• A highly sensitive and specific immunoassay for enrofloxacin was developed.

In the paper, an enzyme-linked immunosorbent immunoassay (ELISA) for detection of enrofloxacin was described using one new derivative of enrofloxacin as coating hapten, resulting in surprisingly high sensitivity and specificity. Incorporation of aminobutyric acid (AA) in the new derivative of enrofloxacin had decreased the IC50 of the ELISA for enrofloxacin from 1.3 μg L−1 to as low as 0.07 μg L−1. The assay showed neglect cross-reactivity for other fluoroquinolones but ofloxacin (8.23%), marbofloxacin (8.97%) and pefloxacin (7.29%). Analysis of enrofloxacin fortified chicken muscle showed average recoveries from 81 to 115%. The high sensitivity and specificity of the assay makes it a suitable screening method for the determination of low levels of enrofloxacin in chicken muscle without clean-up step.

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