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Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Authors :
Roland Zengerle
Thomas Nesch
Melanie Hoehl
Alexander H. Slocum
Nils Paust
Felix von Stetten
Arne Dannenberg
Juergen Steigert
Michael Weißert
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Mechanical Engineering
Slocum, Alexander H.
Hoehl, Melanie M.
Source :
Prof. Slocum via Angie Locknar
Publication Year :
2014
Publisher :
Springer Science and Business Media LLC, 2014.

Abstract

This paper introduces a disposable battery-driven heating system for loop-mediated isothermal DNA amplification (LAMP) inside a centrifugally-driven DNA purification platform (LabTube). We demonstrate LabTube-based fully automated DNA purification of as low as 100 cell-equivalents of verotoxin-producing Escherichia coli (VTEC) in water, milk and apple juice in a laboratory centrifuge, followed by integrated and automated LAMP amplification with a reduction of hands-on time from 45 to 1 min. The heating system consists of two parallel SMD thick film resistors and a NTC as heating and temperature sensing elements. They are driven by a 3 V battery and controlled by a microcontroller. The LAMP reagents are stored in the elution chamber and the amplification starts immediately after the eluate is purged into the chamber. The LabTube, including a microcontroller-based heating system, demonstrates contamination-free and automated sample-to-answer nucleic acid testing within a laboratory centrifuge. The heating system can be easily parallelized within one LabTube and it is deployable for a variety of heating and electrical applications.

Details

ISSN :
15728781 and 13872176
Database :
OpenAIRE
Journal :
Biomedical Microdevices
Accession number :
edsair.doi.dedup.....92365c5e7af48c3c8d1add462331130b
Full Text :
https://doi.org/10.1007/s10544-014-9841-9