Abstract: Antimicrobial susceptibility testing (AST) helps guide clinical rational selection of antibiotics, enabling patients to achieve better prognosis and reducing the development of drug resistance. However, traditional AST is time-consuming and labor-intensive, meaning that clinicians are not able to obtain susceptibility results in a timely manner, and have to rely on experience based on bacterial species and epidemiological data for empirical drug selection. This may be one of the reasons for the continuous increase in the number of drug-resistant strains. In recent years, researchers have focused on the development of rapid and simple AST assays and have made relevant research progress. This article reviews the new AST technologies based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), microfluidic chip systems, nucleic acid detection methods, immunological detection, and flow cytometry, in the hope of providing reference for improving the efficiency of AST testing in clinical microbiology laboratories.