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Anal Chem. 2019 May 7;91(9):6296-6303. doi: 10.1021/acs.analchem.9b01064. Epub 2019 Apr 15.

Rapid Antibiotic Susceptibility Testing of Pathogenic Bacteria Using Heavy-Water-Labeled Single-Cell Raman Spectroscopy in Clinical Samples.

Yang K1,2, Li HZ1,2, Zhu X3, Su JQ1, Ren B4, Zhu YG1,5, Cui L1.

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Key Lab of Urban Environment and Health , Institute of Urban Environment, Chinese Academy of Sciences , Xiamen 361021 , China.
University of Chinese Academy of Sciences , 19A Yuquan Road , Beijing 100049 , China.
The Second Affiliated Hospital of Xiamen Medical College , Xiamen 361021 , China.
Department of Chemistry , Xiamen University , Xiamen 361005 , China.
State Key Laboratory of Urban and Regional Ecology , Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences , Beijing 100085 , China.


Speeding up antibiotic susceptibility testing (AST) is urgently needed in clincial settings to guide fast and tailored antibiotic prescription before treatment. It remains a big challenge to achieve a sample-to-AST answer within a half working day directly from a clinical sample. Here we develop single-cell Raman spectroscopy coupled with heavy water labeling (Raman-D2O) as a rapid activity-based AST approach directly applicable for clinical urine samples. By rapidly transferring (15 min) bacteria in clinical urine for AST, the total assay time from receiving urine to binary susceptibility/resistance (S/R) readout was shortened to only 2.5 h. Moreover, by overcoming the nonsynchronous responses between microbial activity and microbial growth, together with setting a new S/R cutoff value based on relative C-D ratios, S/R of both pathogenic isolates and three clinical urines against antibiotics of different action mechanisms determined by Raman-D2O were all consistent with the slow standard AST assay used in clincial settings. This work promotes clinical practicability and faciliates antibiotic stewardship.

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