NDLI: On-chip parallel detection of foodborne pathogens using loop-mediated isothermal amplification

Content Provider	Springer Nature Link
Author	Duarte, Carlos Salm, Eric Dorvel, Brian Reddy, Bobby Bashir, Rashid
Copyright Year	2013
Abstract	According to estimates issued by the Center for Disease Control and Prevention, one out of six Americans will get sick during this year due to consumption of contaminated products and there will be 50,000 related hospitalizations. To control and treat the responsible foodborne diseases, rapid and accurate detection of pathogens is extremely important. A portable device capable of performing nucleic acid amplification will enable the effective detection of infectious agents in multiple settings, leading to better enforcement of food safety regulations. This work demonstrates the multiplexed detection of food pathogens through loop-mediated isothermal amplification on a silicon chip. Silane passivation is used to prevent the adsorption of the polymerase on silicon oxide, which can severely inhibit nucleic acid amplification. We demonstrate the multiplexed screening of virulence genes of Listeria monocytogenes, Escherichia coli, and Salmonella by dehydrating the corresponding primers in oxidized silicon wells. Droplets of 30 nL with reagents for nucleic acid amplification and lysate of suspected pathogens are arrayed on micro-machined wells with an automated microinjection system. We show that dehydrated primers re-suspend when other reagents are microinjected, and the resulting mix can be used to specifically amplify the targeted gene. Results of characterization experiments demonstrate sensitivity down to a few templates per reaction, specificity that enables multiplexed screening, and robustness that allows amplification without DNA extraction.
Starting Page	821
Ending Page	830
Page Count	10
File Format	PDF
ISSN	13872176
Journal	Biomedical Microdevices
Volume Number	15
Issue Number	5
e-ISSN	15728781
Language	English
Publisher	Springer US
Publisher Date	2013-04-26
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Miniaturized DNA amplification Loop-mediated isothermal amplification Primer dehydration Silane passivation Multiplexed screening Biomedical Engineering Biophysics and Biological Physics Nanotechnology Engineering Fluid Dynamics
Content Type	Text
Resource Type	Article
Subject	Nanoscience and Nanotechnology Molecular Biology Biomedical Engineering

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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