Single cell antimicrobial susceptibility testing by confined microchannels and electrokinetic loading.
Single cell antimicrobial susceptibility testing by confined microchannels and electrokinetic loading.
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DOI：
10.1021/ac4004248
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发表时间：
2013-04-16
影响因子：
7.4
通讯作者：
Wong, Pak Kin
中科院分区：
文献类型：
--
作者：
Lu, Yi;Gao, Jian;Zhang, Donna D.;Gau, Vincent;Liao, Joseph C.;Wong, Pak Kin
多重耐药病原体是一个新出现的全球健康问题。除了需要开发新的抗生素外，快速确定细菌抗生素耐药性的能力是管理传染病和预防多重耐药病原体的最关键步骤之一。在这里，我们报告了一个单细胞抗菌药物敏感性测试（AST）的方法，用于快速确定细菌病原体的抗生素耐药性。通过将单个细菌限制在具有与单个细菌相当的尺寸的透气微通道中，可以在单细胞水平上实时监测细菌的抗生素抗性。为了便于细菌动态加载到封闭的微通道中进行观察，AC电动力学被证明用于将细菌捕获到高电导率AST缓冲液中的限定位置。电动技术实现了约75%的负载效率，对细菌生长速率的影响可以忽略不计。为了优化单细胞AST的方案，系统地测定了不同抗生素条件下单个细菌的细菌生长速率。通过快速测定Mueller-Hinton培养基和尿液中尿路致病性临床分离株的抗菌药物耐药性谱，证明了单细胞AST的适用性。细菌的抗生素耐药性谱可以在不到一小时内确定，而标准的基于培养的AST技术需要几天。
Multidrug-resistant pathogens are an emerging global health problem. In addition to the need of developing new antibiotics in the pipeline, the ability to rapidly determine the antibiotic resistance profiles of bacteria represents one of the most crucial steps toward the management of infectious diseases and the prevention of multidrug-resistant pathogens. Here, we report a single cell antimicrobial susceptibility testing (AST) approach for rapid determination of the antibiotic resistance of bacterial pathogens. By confining individual bacteria in gas permeable microchannels with dimensions comparable to a single bacterium, the antibiotic resistance of the bacteria can be monitored in real-time at the single cell level. To facilitate the dynamic loading of the bacteria into the confined microchannels for observation, AC electrokinetics is demonstrated for capturing bacteria to defined locations in high-conductivity AST buffer. The electrokinetic technique achieves a loading efficiency of about 75% with a negligible effect on the bacterial growth rate. To optimize the protocol for single cell AST, the bacterial growth rate of individual bacteria under different antibiotic conditions has been determined systematically. The applicability of single cell AST is demonstrated by the rapid determination of the antimicrobial resistant profiles of uropathogenic clinical isolates in Mueller-Hinton media and in urine. The antibiotic resistance profiles of bacteria can be determined in less than one hour compared to days in standard culture-based AST techniques.
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影响因子：
13.8
作者：
Mach KE;Wong PK;Liao JC
通讯作者：
Liao JC
影响因子：
6.4
作者：
Daum, RS;Ito, T;Boyle-Wang, S
通讯作者：
Boyle-Wang, S
影响因子：
7.4
作者：
Mann, Thomas S.;Mikkelsen, Susan R.
通讯作者：
Mikkelsen, Susan R.
影响因子：
7.4
作者：
Ertl, P;Robello, E;Mikkelsen, SR
通讯作者：
Mikkelsen, SR
影响因子：
6.1
作者：
Peitz, Ingmar;van Leeuwen, Rien
通讯作者：
van Leeuwen, Rien