As an important member of tool enzymes, exonuclease is a kind of hydrolytic enzymes without strict base sequence dependent. In recent years, by taking advantage of different hydrolysis ways of exonuclease and nanotechnology, cycle effect of enzyme digestion, aptamer, non Watson-Crick base pairing system by metal ions, fluorescent nucleic acid probes, electrochemical methods etc. , a series of exonuclease-assisted signal amplification strategies have been developed, which played a very key role in improving the sensitivity of detection methods. Therefore, exonuclease has been widely used in high sensitive detection of nucleic acids, proteins, ions, small molecules and so on. To understand it better and apply it well in the future, the application progress of exonuclease-assisted signal amplification strategies in biochemical analysis has been summarized in this review.