The invention provides an enzyme-linked immunoassay kit for detecting a fluoroquinolone medicine. The enzyme-linked immunoassay kit comprises an enzyme-labelled plate coated with a coating antigen, a standard solution of the fluoroquinolone medicine, an enzyme-labelled bivalent antibody, a specific antibody of the fluoroquinolone medicine, a developing solution of a substrate, a stop solution, a washing solution, a combination solution and a diluent of a honey sample; the coating antigen is a coupled antigen of the fluoroquinolone medicine; and the enzyme-labelled bivalent antibody is an enzyme-labelled anti-goat and anti-rat antibody. The invention further discloses a method for detecting the fluoroquinolone medicine by using the enzyme-linked immunoassay kit. The method comprises the following steps: pre-processing a sample at first, then, detecting by using the kit, and finally, analyzing a detection result. The enzyme-linked immunoassay kit provided by the invention can be used for detecting the content of the fluoroquinolone medicine in honey, chicken, pork, fish, shrimp, milk, milk powder and egg samples, is simple and convenient to operate, low in cost and high in sensitivity and can be used for monitoring on site and applied to screening of a lot of samples.