使用无膜微流控透析和介电泳从血液中分离和浓缩细菌。
Isolation and concentration of bacteria from blood using microfluidic membraneless dialysis and dielectrophoresis.
机构信息
Alkek Center for Metagenomics and Microbiome Research, Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA.
Department of Infection Control and Employee Health, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
出版信息
Lab Chip. 2017 Mar 29;17(7):1340-1348. doi: 10.1039/c6lc01277a.
A microfluidic system that combines membraneless microfluidic dialysis and dielectrophoresis to achieve label-free isolation and concentration of bacteria from whole blood is presented. Target bacteria and undesired blood cells are discriminated on the basis of their differential susceptibility to permeabilizing agents that alter the dielectrophoretic behavior of blood cells but not bacteria. The combined membraneless microdialysis and dielectrophoresis system isolated 79 ± 3% of Escherichia coli and 78 ± 2% of Staphylococcus aureus spiked into whole blood at a processing rate of 0.6 mL h. Collection efficiency was independent of the number of target bacteria up to 10 cells. Quantitative PCR analysis revealed that bacterial 16S rDNA levels were enriched more than 307-fold over human DNA in the fraction recovered from the isolation system compared with the original specimen. These data demonstrate feasibility for an instrument to accelerate the detection and analysis of bacteria in blood by first isolating and concentrating them in a microchamber.
本文提出了一种将无膜微流控透析和介电泳相结合的微流控系统，可实现对全血中细菌的无标记分离和浓缩。基于对改变血细胞介电泳行为但不改变细菌介电泳行为的通透性试剂的不同敏感性，对靶细菌和不需要的血细胞进行区分。组合的无膜微透析和介电泳系统可从全血中分离出 79±3%的大肠杆菌和 78±2%的金黄色葡萄球菌，处理速度为 0.6mL/h。收集效率与目标细菌数量无关，最高可达 10 个细胞。定量 PCR 分析显示，与原始标本相比，从分离系统中回收的部分细菌 16S rDNA 水平比人 DNA 富集了 307 倍以上。这些数据证明了一种仪器的可行性，该仪器可以通过首先将它们在微腔室内分离和浓缩来加速血液中细菌的检测和分析。
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