A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections.
A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections.
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DOI：
10.3390/bios11080288
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发表时间：
2021-08-22
期刊：
影响因子：
--
通讯作者：
Wong PK
中科院分区：
文献类型：
--
作者：
Forsyth B;Torab P;Lee JH;Malcom T;Wang TH;Liao JC;Yang S;Kvam E;Puleo C;Wong PK
血流感染是世界范围内发病率和死亡率的重要原因。快速开始有效的抗生素治疗对血流感染患者至关重要。然而，血液传播病原体的诊断在很大程度上由于血液的基质效应和漫长的试管培养程序而变得复杂。在这里，我们报告了一种用于从全血中快速分离和富集细菌病原体的无培养工作流程，用于单细胞抗菌药物敏感性测试（AST）。葡聚糖沉降步骤在20-30分钟内将血细胞浓度降低4个数量级，同时保持样品中细菌的有效浓度。红细胞去除有助于在脓毒症相关细菌浓度下进行下游基于离心的富集步骤。该工作流程与常见的耐药细菌兼容，不影响最低抑菌浓度。通过应用微流控单细胞捕获装置，我们展示了在单细胞水平上快速确定细菌感染和抗菌药物敏感性测试的工作流程。从血液到分类AST结果的整个工作流程可以在不到两个小时内完成。
Bloodstream infections are a significant cause of morbidity and mortality worldwide. The rapid initiation of effective antibiotic treatment is critical for patients with bloodstream infections. However, the diagnosis of bloodborne pathogens is largely complicated by the matrix effect of blood and the lengthy blood tube culture procedure. Here we report a culture-free workflow for the rapid isolation and enrichment of bacterial pathogens from whole blood for single-cell antimicrobial susceptibility testing (AST). A dextran sedimentation step reduces the concentration of blood cells by 4 orders of magnitude in 20–30 min while maintaining the effective concentration of bacteria in the sample. Red blood cell depletion facilitates the downstream centrifugation-based enrichment step at a sepsis-relevant bacteria concentration. The workflow is compatible with common antibiotic-resistant bacteria and does not influence the minimum inhibitory concentrations. By applying a microfluidic single-cell trapping device, we demonstrate the workflow for the rapid determination of bacterial infection and antimicrobial susceptibility testing at the single-cell level. The entire workflow from blood to categorical AST result can be completed in less than two hours.
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影响因子：
7.4
作者：
Lu, Yi;Gao, Jian;Zhang, Donna D.;Gau, Vincent;Liao, Joseph C.;Wong, Pak Kin
通讯作者：
Wong, Pak Kin
影响因子：
4.9
作者：
Diekema, Daniel J.;Hsueh, Po-Ren;Jones, Ronald N.
通讯作者：
Jones, Ronald N.
影响因子：
5.3
作者：
Pinkofsky, HB
通讯作者：
Pinkofsky, HB
影响因子：
--
作者：
Kadlec, Meichei Wang;You, David;Wong, Pak Kin
通讯作者：
Wong, Pak Kin
DOI：
10.1038/nrurol.2017.20
发表时间：
2017-05
期刊：
Nature reviews. Urology
影响因子：
--
作者：
Davenport M;Mach KE;Shortliffe LMD;Banaei N;Wang TH;Liao JC
通讯作者：
Liao JC