The invention provides an enzyme linked immune regent box to detect gentamycin. It includes: the enzyme labeled board which encrusts the encrusting substance, the enzyme marker, the special antibody for the gentamycin (when the antigen is on enzyme labeled board and the enzyme marker is the enzyme labeled antiantibody or the antiantibody is in the enzyme labeled board and the enzyme marker is theenzyme labeled antigen), the gentamycin standard solution, the substrate color solution, the stop solution, the condense washing solution, the condense recovery solution. The invention also disclosesthe method which includes the process: first to pre-treat the sample, then to detect by the regent box and last to analyze the detecting result. The invention is to detect the gentamycin residue weight in the animal food such as the chicken, chicken liver, pork, pork liver, milk powder, egg and feedstuff. The method is simple, low cost and has high sensitivity, also it is proper for detect the mass sample.