Abstract: Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS) has achieved remarkable progress in diagnosis of infectious diseases in recent years, showing certain advantages in identification of bacteria and fungi as well as analysis of macromolecules such as proteins.The application effectiveness of MALDI-TOF mass spectrometry in detection of infectious diseases, especially foodborne pathogenic bacteria infections, urinary tract infections, bloodstream infections, reproductive system infections as well as the drug resistance testing, was reviewed in the article so as to facilitate the diagnosis and treatment of infectious diseases.