Isolation and concentration of bacteria from blood using microfluidic membraneless dialysis and dielectrophoresis

L. D'Amico; N. J. Ajami; J. A. Adachi; P. R.C. Gascoyne; J. F. Petrosino

doi:10.1039/c6lc01277a

Isolation and concentration of bacteria from blood using microfluidic membraneless dialysis and dielectrophoresis

L. D'Amico, N. J. Ajami, J. A. Adachi, P. R.C. Gascoyne, J. F. Petrosino

Infectious Diseases, Infection Control, and Employee Health

Research output: Contribution to journal › Article › peer-review

59 Scopus citations

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 10⁵ 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.

Original language	English (US)
Pages (from-to)	1340-1348
Number of pages	9
Journal	Lab on a Chip
Volume	17
Issue number	7
DOIs	https://doi.org/10.1039/c6lc01277a
State	Published - Apr 7 2017

ASJC Scopus subject areas

Bioengineering
Biochemistry
General Chemistry
Biomedical Engineering

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10.1039/c6lc01277a

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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 105 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.",

author = "L. D'Amico and Ajami, {N. J.} and Adachi, {J. A.} and Gascoyne, {P. R.C.} and Petrosino, {J. F.}",

note = "Funding Information: The authors would like to thank Jeffery Tarrand, MD, for sharing his knowledge of clinical microbiology, and Thomas Anderson for helping design the microfluidic devices. The quadrupole microelectrodes were gifted by Hywel Morgan, PhD and Nicolas Green, PhD, from the Centre for Hybrid Biodevices at the University of Southampton. Funding was provided by the Cancer Prevention Research Institute of Texas (RP130476) and the Baylor College of Medicine Alkek Center for Metagenomics and Microbiome Research. LD was supported in part by the Whitaker Foundation Summer Research grant and the National Institute of Biomedical Imaging and Bioengineering Imaging Sciences training grant (grant number 1 T32 EB 7507-1 A2). Publisher Copyright: {\textcopyright} The Royal Society of Chemistry.",

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AU - Gascoyne, P. R.C.

AU - Petrosino, J. F.

N1 - Funding Information: The authors would like to thank Jeffery Tarrand, MD, for sharing his knowledge of clinical microbiology, and Thomas Anderson for helping design the microfluidic devices. The quadrupole microelectrodes were gifted by Hywel Morgan, PhD and Nicolas Green, PhD, from the Centre for Hybrid Biodevices at the University of Southampton. Funding was provided by the Cancer Prevention Research Institute of Texas (RP130476) and the Baylor College of Medicine Alkek Center for Metagenomics and Microbiome Research. LD was supported in part by the Whitaker Foundation Summer Research grant and the National Institute of Biomedical Imaging and Bioengineering Imaging Sciences training grant (grant number 1 T32 EB 7507-1 A2). Publisher Copyright: © The Royal Society of Chemistry.

PY - 2017/4/7

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N2 - 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 105 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.

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Isolation and concentration of bacteria from blood using microfluidic membraneless dialysis and dielectrophoresis

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