Abstract: In the global agricultural production process, the irregular use of chemical pesticides has led to excessive pesticide residues in agricultural products, posing a potential threat to human health. In order to effectively monitor pesticide residues, enzyme-linked immunosorbent assay (ELISA) with high throughput, simple operation and low cost is commonly used for rapid detection of pesticide residues. However, the sensitivity of traditional ELISA cannot meet the requirements of pesticide residue detection. To address this issue, researchers have developed a variety of strategies to improve the sensitivity of ELISA, among which the enhancement of molecular element recognition and the improvement of signal transduction efficiency have attracted widespread attention in enhanced immunoassay. This article provides a detailed introduction to the improvement of ELISA sensitivity by high-affinity antibodies, low-affinity antigens, increased enzyme loading, high-performance nanozymes and enzyme cascade reactions. The challenges and development directions of the future application of ELISA in pesticide residue detection were discussed, providing reference for high-sensitivity rapid detection technology of pesticide residues.