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Rapid antimicrobial susceptibility testing (AST) is urgently needed fortreating infections with correct antibiotics and slowing down the emergence ofantibiotic-resistant bacteria. Here, we report a phenotypic platform thatrapidly produces AST results by femtosecond stimulated Raman scattering imagingof deuterium oxide (D2O) metabolism. Metabolic incorporation of D2O intobiomass in a single bacterium is probed in as short as 10 minutes after culturein 70% D2O medium, the fastest among current technologies. Single-cellmetabolism inactivation concentration (SC-MIC) is obtained in less than 2.5hours from colony to results. The SC-MIC results of 37 sets of samples, whichinclude 8 major bacterial species and 14 different antibiotics oftenencountered in clinic, are validated by standard minimal inhibitoryconcentration blindly measured via broth microdilution. Towards clinicaltranslation, SRS imaging of D2O metabolic incorporation and SC-MICdetermination after 1-h antibiotics treatment and 30-minutes mixture of D2O andantibiotics incubation of bacteria in urine or whole blood is demonstrated.

Rapid Determination of Antimicrobial Susceptibility by Stimulated Raman Scattering Imaging of D2O Metabolic Incorporation in a Single Bacterium