Detecting bacteria and determining their susceptibility to antibiotics by stochastic confinement in nanoliter droplets using plug-based microfluidics.
Detecting bacteria and determining their susceptibility to antibiotics by stochastic confinement in nanoliter droplets using plug-based microfluidics.
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使用基于插头的微流体剂在纳米液液滴中检测细菌并确定其对抗生素的敏感性。
DOI：
10.1039/b804911d
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发表时间：
2008-08
期刊：
影响因子：
6.1
通讯作者：
Ismagilov RF
中科院分区：
文献类型：
--
作者：
Boedicker JQ;Li L;Kline TR;Ismagilov RF
本文介绍了基于插头的微流控技术，该技术能够快速检测和筛选样本中的细菌，包括复杂的生物基质，而不需要预先培养。与传统的细菌培养和检测方法不同，传统的细菌培养和检测方法依赖于孵育样本来将细菌浓度增加到可检测到的水平，这种方法将单个细菌限制在体积上的液滴纳米升中。当单个细胞被限制在小体积的塞子中时，每个塞子的负载少于一个细菌，检测时间与塞子体积成正比。限制增加了细胞密度，允许释放的分子在细胞周围积累，省去了预孵化步骤，减少了检测细菌所需的时间。我们把这种方法称为“随机禁闭”。采用微流控杂交法，一次性测定耐甲氧西林金黄色葡萄球菌(MRSA)对多种抗生素的药敏图谱，并测定头孢西丁(CFX)对该菌株的最低抑菌浓度(MIC)。此外，该技术还被用于区分人血浆样本中的金黄色葡萄球菌敏感株和耐药株。高通量微流控技术与单细胞测量相结合，还可以在含有细菌的单个样本上同时进行多项测试。这项技术可以提供一种快速有效的针对患者的细菌感染治疗方法，并可以扩展到需要在更短的时间内对细菌样本进行多种功能测试的各种应用。
This article describes plug-based microfluidic technology that enables rapid detection and drug susceptibility screening of bacteria in samples, including complex biological matrices, without pre-incubation. Unlike conventional bacterial culture and detection methods, which rely on incubation of a sample to increase the concentration of bacteria to detectable levels, this method confines individual bacteria into droplets nanoliters in volume. When single cells are confined into plugs of small volume such that the loading is less than one bacterium per plug, the detection time is proportional to plug volume. Confinement increases cell density and allows released molecules to accumulate around the cell, eliminating the pre-incubation step and reducing the time required to detect the bacteria. We refer to this approach as ‘stochastic confinement’. Using the microfluidic hybrid method, this technology was used to determine the antibiogram — or chart of antibiotic sensitivity — of methicillin-resistant Staphylococcus aureus (MRSA) to many antibiotics in a single experiment and to measure the minimal inhibitory concentration (MIC) of the drug cefoxitin (CFX) against this strain. In addition, this technology was used to distinguish between sensitive and resistant strains of S. aureus in samples of humanblood plasma. High-throughput microfluidic techniques combined with single-cell measurements also enable multiple tests to be performed simultaneously on a single sample containing bacteria. This technology may provide a method of rapid and effective patient-specific treatment of bacterial infections and could be extended to a variety of applications that require multiple functional tests of bacterial samples on reduced timescales.
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作者：
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通讯作者：
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影响因子：
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