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

• Published in: Analytical chemistry (Washington) 2019-05, Vol.91 (9), p.6296-6303
• Main Authors: Yang, Kai, Li, Hong-Zhe, Zhu, Xuan, Su, Jian-Qiang, Ren, Bin, Zhu, Yong-Guan, Cui, Li
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Antibiotics; Bacteria; Biological activity; Chemistry; Heavy water; Microbial activity; Microorganisms; Raman spectroscopy; Spectroscopy; Spectrum analysis; Urine
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.9b01064

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.