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Detecting bacteria and determining their susceptibility to antibiotics by stochastic confinement in nanoliter droplets using plug-based microfluidics

Boedicker, James Q. and Li, Liang and Kline, Timothy R. and Ismagilov, Rustem F. (2008) Detecting bacteria and determining their susceptibility to antibiotics by stochastic confinement in nanoliter droplets using plug-based microfluidics. Lab on a Chip, 8 (8). pp. 1265-1272. ISSN 1473-0197. doi:10.1039/B804911D. https://resolver.caltech.edu/CaltechAUTHORS:20130821-160716197

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Abstract

This article describes plug-based microfluidic technology that enables rapid detection and drug susceptibility screening of bacteria in samples, including complex biological matrices, without preincubation. 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 human blood 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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1039/B804911DDOIArticle
http://pubs.rsc.org/en/Content/ArticleLanding/2008/LC/b804911dPublisherArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2612531/PubMed CentralArticle
http://www.rsc.org/suppdata/lc/b8/b804911d/b804911d.pdfPublisherSupporting Information
ORCID:
AuthorORCID
Ismagilov, Rustem F.0000-0002-3680-4399
Additional Information:This journal is © The Royal Society of Chemistry 2008. Received 26th March 2008, Accepted 21st May 2008. First published on the web 4th July 2008. This work was supported in part by the NSF CRC (Grant No. 0526693) and by the NIH. We would like to thank Christian Kastrup, Helen Song, Urs Spitz, and Lukas Wick for helpful comments and Jessica M. Price for contributions in writing and editing this manuscript. R. F. I. is a Cottrell Scholar of Research Corporation and an A. P. Sloan Research Fellow. Some of this work was performed at the Materials Research Science and Engineering Centers microfluidic facility funded by the NSF.
Funders:
Funding AgencyGrant Number
NSF0526693
NIHUNSPECIFIED
Cottrell Scholar of Research CorporationUNSPECIFIED
Alfred P. Sloan FoundationUNSPECIFIED
Issue or Number:8
DOI:10.1039/B804911D
Record Number:CaltechAUTHORS:20130821-160716197
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20130821-160716197
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:40778
Collection:CaltechAUTHORS
Deposited By: Whitney Barlow
Deposited On:27 Aug 2013 17:47
Last Modified:10 Nov 2021 00:08

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