Abstract: As a microorganism identification system, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been developed as a mature technique system. MALDI-TOF MS has shown great potential in the field of pathogen resistance analysis due to its simplicity, rapidity, cost-effectiveness, high throughput, high sensitivity and specificity. In addition to the wide use in bacterial identification, the researches on antibiotic resistance of pathogens based on MALDI-TOF MS have made great progress in recent years. This paper summarizes the progress in research of the application of MALDI-TOF MS in pathogen antibiotic resistance analysis from qualitative and quantitative perspectives and descripts the application prospect of this technique.