Abstract
Blood-stream infections (BSI) remain a major health challenge, with an increasing incidence worldwide and a high mortality rate. Early treatment with appropriate antibiotics can reduce BSI-related morbidity and mortality, but success requires rapid identification of the infecting organisms. The rapid, culture-independent diagnosis of BSI could be significantly facilitated by straightforward isolation of highly purified bacteria from whole blood. We present a microfluidic-based, sample-preparation system that rapidly and selectively lyses all blood cells while it extracts intact bacteria for downstream analysis. Whole blood is exposed to a mild detergent, which lyses most blood cells, and then to osmotic shock using deionized water, which eliminates the remaining white blood cells. The recovered bacteria are 100 % viable, which opens up possibilities for performing drug susceptibility tests and for nucleic-acid-based molecular identification.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Hansson J, Karlsson JM, Haraldsson T, Brismar H, van der Wijngaart W, Russom A (2012) Inertial microfluidics in parallel channels for high-throughput applications. Lab Chip 12:4644–4650
Kumar A, Roberts D, Wood KE, Light B, Parrillo JE, Sharma S, Suppes R, Feinstein D, Zanotti S, Taiberg L, Gurka D, Kumar A, Cheang M (2006) Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 34:1589–1596
Lee JJ, Jeong KJ, Hashimoto M, Kwon AH, Rwei A, Shankarappa SA, Tsui JH, Kohane DS (2014) Synthetic ligand-coated magnetic nanoparticles for microfluidic bacterial separation from blood. Nano Lett 14:1–5
Mach AJ, Di Carlo D (2010) Continuous scalable blood filtration device using inertial microfluidics. Biotechnol Bioeng 107:302–311
Mariella R Jr (2008) Sample preparation: the weak link in microfluidics-based biodetection. Biomed Microdev 10:777–784
Mohan R, Mukherjee A, Sevgen SE, Sanpitakseree C, Lee J, Schroeder CM, Kenis PJ (2013) A multiplexed microfluidic platform for rapid antibiotic susceptibility testing. Biosens Bioelectron 49:118–125
Park S, Zhang Y, Wang TH, Yang S (2011) Continuous dielectrophoretic bacterial separation and concentration from physiological media of high conductivity. Lab Chip 11:2893–2900
Tang Y, Zhen L, Liu J, Wu J (2013) Rapid antibiotic susceptibility testing in a microfluidic pH sensor. Anal Chem 85:2787–2794
Wang S, Inci F, Chaunzwa TL, Ramanujam A, Vasudevan A, Subramanian S, Chi Fai Ip A, Sridharan B, Gurkan UA, Demirci U (2012) Portable microfluidic chip for detection of Escherichia coli in produce and blood. Int J Nanomed 7:2591–2600
Wu Z, Willing B, Bjerketorp J, Jansson JK, Hjort K (2009) Soft inertial microfluidics for high throughput separation of bacteria from human blood cells. Lab Chip 9:1193–1199
Xia Y, Whitesides GM (1998) Soft lithography. Angew Chem Int Ed Engl 37:550–575
Acknowledgments
This work was sponsored by the European Commission through the projects FP7 InTopSens and IMI RAPP-ID.
Supporting information
Supplementary Figure 1—Experimental design of the selective cell lysis protocol on a macro scale.
Supplementary Figure 2—Selection of the most effective detergent and the optimal detergent concentration in macro scale experiments.
Supplementary Figure 3—Titration of the bacteria concentration spiked in blood or PBS in 1 % saponin-based lysis solution.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Zelenin, S., Hansson, J., Ardabili, S. et al. Microfluidic-based isolation of bacteria from whole blood for sepsis diagnostics. Biotechnol Lett 37, 825–830 (2015). https://doi.org/10.1007/s10529-014-1734-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-014-1734-8