The invention discloses a method for rapid detection of drug resistance phenotypes of strains. The method includes: firstly, preparing antibacterial drug diluent in gradient-changing concentration from nutrient broth, dropwise adding antibacterial drugs in different concentrates onto sample target plates of a mass spectrometer to form drug-sensitive target plates, and drying for standby application; secondly, preparing standard bacterial suspension of to-be-tested strain from sterile saline solution, and diluting with nutrient broth to keep for standby application; thirdly, sequentially dropwise adding the diluted standard bacterial suspension onto the prepared drug-sensitive target plates to form microdroplets, putting into a wet box, incubating for 2h-6h in an incubator, and discarding broth on the drug-sensitive target plates; fourthly, after drying, dropwise adding a mass spectrometry pretreatment agent, crystallizing, performing microbiological assay by adopting a mass spectrometer, and reading a result. The method is simple in operation, low in material consumption and independent from any drug resistance mechanisms, antibiotic coating and concentrations can be determined according to demands, quickness in detection is achieved, the result is direct and clear, excessive analysis is avoided, and the method is wide in application range.