D2O代谢掺入的受激拉曼散射成像快速测定细菌的抗菌药物敏感性
Rapid Determination of Antimicrobial Susceptibility by Stimulated Raman Scattering Imaging of D2O Metabolic Incorporation in a Single Bacterium
摘要：
快速抗菌药物敏感性测试（AST）是迫切需要正确的抗生素治疗感染和减缓耐药性细菌的出现。在这里，我们报告了一个表型平台，通过氧化氘（D2 O）代谢的飞秒受激拉曼散射成像快速产生AST结果。在70%D2O培养基中培养后，在单个细菌中将D2 O代谢掺入生物质中的时间短至10分钟，这是目前技术中最快的。单细胞代谢灭活浓度（SC-MIC）从菌落到结果在2.5小时内获得。采用微量肉汤稀释法盲测标准最低抑菌浓度，对临床常见的8种主要细菌和14种抗生素共37组样品的SC-MIC结果进行验证。对于临床翻译，D2 O代谢掺入和SC-MIC测定后1小时的抗生素治疗和30分钟的混合物的D2 O和抗生素孵育的细菌在尿液或全血中的SRS成像证明。
英文摘要：
Rapid antimicrobial susceptibility testing (AST) is urgently needed for treating infections with correct antibiotics and slowing down the emergence of antibiotic-resistant bacteria. Here, we report a phenotypic platform that rapidly produces AST results by femtosecond stimulated Raman scattering imaging of deuterium oxide (D2O) metabolism. Metabolic incorporation of D2O into biomass in a single bacterium is probed in as short as 10 minutes after culture in 70% D2O medium, the fastest among current technologies. Single-cell metabolism inactivation concentration (SC-MIC) is obtained in less than 2.5 hours from colony to results. The SC-MIC results of 37 sets of samples, which include 8 major bacterial species and 14 different antibiotics often encountered in clinic, are validated by standard minimal inhibitory concentration blindly measured via broth microdilution. Towards clinical translation, SRS imaging of D2O metabolic incorporation and SC-MIC determination after 1-h antibiotics treatment and 30-minutes mixture of D2O and antibiotics incubation of bacteria in urine or whole blood is demonstrated.
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