Objective To develop a novel ELISA for sensitive and naked-eye detection of danofloxacin （DAN）.Methods Based on TYR signal amplification， a novel ELISA was developed for sensitive detection of DAN. Analytical performance of the method was conducted by sensitivity and specificity experiments. The method was used for detecting DAN-spiked chicken and pork samples to verify.Results This method could be used to qualitatively detect DAN with naked eyes at a cut-off value of 0.4 ng/mL， which was 25-fold lower than that of the traditional ELISA （10 ng/mL）. The method exhibited no cross-reaction of DAN with six other drugs in buffer. The method was successfully used for detecting DAN-spiked chicken and pork samples.Conclusion The developed novel ELISA exhibits high sensitivity and specificity， and can be used for the danofloxacin detection in actual samples by naked eye.