title: Isolation and concentration of bacteria from blood using microfluidic membraneless dialysis and dielectrophoresis.
abstract: 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-1. Collection efficiency was independent of the number of target bacteria up to 105cells. 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.
authors.affiliation: Alkek Center for Metagenomics and Microbiome Research, Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA. jpetrosi@bcm.edu and Advanced Electrofluidic Systems, LLC, Houston, Texas, USA.
authors.forename: L
authors.identifier: 0000-0001-9839-7568
authors.initials: L
authors.lastname: D'Amico
authors.affiliation: Alkek Center for Metagenomics and Microbiome Research, Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA. jpetrosi@bcm.edu.
authors.forename: N J
authors.initials: NJ
authors.lastname: Ajami
authors.affiliation: Department of Infection Control and Employee Health, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Adachi
authors.affiliation: Department of Imaging Physics, University of Texas MD Anderson Cancer Center, Houston, TX, USA and Department of Biomedical Engineering, University of Texas at Austin, Austin, TX, USA and Advanced Electrofluidic Systems, LLC, Houston, Texas, USA.
authors.forename: P R C
authors.initials: PR
authors.lastname: Gascoyne
authors.affiliation: Alkek Center for Metagenomics and Microbiome Research, Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA. jpetrosi@bcm.edu.
authors.forename: J F
authors.initials: JF
authors.lastname: Petrosino
mesh_terms: D016470:Bacteremia / Q000175:diagnosis / Q000382:microbiology; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D058491:Bacterial Load / Q000379:methods*; D003198:Computer Simulation; D004269:DNA, Bacterial / Q000032:analysis; D004586:Electrophoresis / Q000295:instrumentation*; D004867:Equipment Design; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D012336:RNA, Ribosomal, 16S / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S