Rapid antimicrobial susceptibility testing (AST) is urgently needed for informing treatment decisions and preventing the spread of antimicrobial resistance resulting from the misuse and overuse of antibiotics. To date, no phenotypic AST exists that can be performed within a single patient visit (30 min) directly from clinical samples. We show that AST results can be obtained by using digital nucleic acid quantification to measure the phenotypic response of Escherichia coli present within clinical urine samples exposed to an antibiotic for 15 min. We performed this rapid AST using our ultrafast (~7 min) digital real-time loop-mediated isothermal amplification (dLAMP) assay [area under the curve (AUC), 0.96] and compared the results to a commercial (~2 hours) digital polymerase chain reaction assay (AUC, 0.98). The rapid dLAMP assay can be used with SlipChip microfluidic devices to determine the phenotypic antibiotic susceptibility of E. coli directly from clinical urine samples in less than 30 min. With further development for additional pathogens, antibiotics, and sample types, rapid digital AST (dAST) could enable rapid clinical decision-making, improve management of infectious diseases, and facilitate antimicrobial stewardship.

迫切需要快速的抗菌药物敏感性测试（AST），以告知治疗决策并防止因滥用和滥用抗生素而引起的抗菌素耐药性扩散。迄今为止，尚无可直接从临床样本中进行单次患者就诊（30分钟）内进行的表型AST。我们表明，可以通过使用数字核酸定量来测量大肠杆菌的表型反应来获得AST结果。在暴露于抗生素15分钟的临床尿液样本中存在。我们使用超快（〜7分钟）数字实时回路介导的等温扩增（dLAMP）分析[曲线下面积（AUC），0.96]进行了这种快速AST，并将结果与​​商用（〜2小时）数字聚合酶链反应分析（AUC，0.98）。快速dLAMP分析可与SlipChip微流控设备一起使用，以在不到30分钟的时间内直接从临床尿液样本中确定大肠杆菌的表型抗生素敏感性。随着对其他​​病原体，抗生素和样品类型的进一步开发，快速数字AST（dAST）可以实现快速的临床决策，改善传染病的管理并促进抗菌素的管理。