Antibiotics, as animal therapeutic agents and growth promoters, are widely used in agriculture, animal husbandry and aquatic product breeding, resulting in excessive antibiotic residues in animal and plant food, which seriously threaten human health. Therefore, detection of antibiotic residues in food is of great significance. However, the existing detection methods of antibiotic residues, such as microbiological assay, enzyme-linked immunoassay, liquid chromatography-mass spectrometry, capillary electrophoresis, usually have the disadvantages of time consuming, complex operation and high cost. As a new technology, biosensor has the advantages of being rapid and simple, high sensitivity, good selectivity and low cost, indicating great potential in the field of antibiotic residue detection. Nucleic acid probe, as a new tool for biological analysis, has been widely used in the development of biosensors. The introduction of nucleic acid probe to the biosensing detection of antibiotic residues opens up a new way for the efficient detection of antibiotic residues. The application of nucleic acid probe in the biosensing detection of antibiotic residue is reviewed from the aspects of electrochemical biosensor, fluorescence biosensor, colorimetric biosensor and other common biosensors, and the future development prospect of this field is also prospected.