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

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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.

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Abbreviations

E. coli :

Escherichia coli

LAMP:

Loop-mediated isothermal DNA amplification

VTEC:

Verotoxin producing E. coli

DNA yield:

Extracted DNA copies, which here are quantified by qPCR

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Acknowledgments

We thank the Harvard-MIT Division of Health Sciences and Technology, as well as the Legatum Center at MIT for support. Thanks to the CR/ARY2 team at Robert Bosch GmbH, particularly F. Laermer, B. Faltin, J. Hoffmann and K. Lemuth. We thank the staff at Nesch Engineering for the productive collaboration, and H.-E.Manneck from Mast Diagnostica for technical support.

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Authors and Affiliations

  1. Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA

    Melanie M. Hoehl & Alexander H. Slocum

  2. Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Melanie M. Hoehl & Alexander H. Slocum

  3. Advanced Research and Engineering, Robert Bosch GmbH, Gerlingen, Germany

    Melanie M. Hoehl, Michael Weißert, Arne Dannenberg & Juergen Steigert

  4. Microsystems Technology, University of Stuttgart, Stuttgart, Germany

    Michael Weißert

  5. Nesch Engineering UG (haftungsbeschränkt), Haigerloch, Germany

    Thomas Nesch

  6. HSG-IMIT, Freiburg, Germany

    Nils Paust, Felix von Stetten & Roland Zengerle

Authors
  1. Melanie M. Hoehl
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  2. Michael Weißert
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  3. Arne Dannenberg
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  4. Thomas Nesch
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  7. Roland Zengerle
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  8. Alexander H. Slocum
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  9. Juergen Steigert
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Corresponding authors

Correspondence to Melanie M. Hoehl or Alexander H. Slocum.

Additional information

Alexander H. Slocum and Juergen Steigert contributed equally to this work.

Electronic supplementary material

The ESI contains details on the LAMP stability testing, the power supply, heater selection and control, as well as LabTube extraction of whole E.coli cells.

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Hoehl, M.M., Weißert, M., Dannenberg, A. et al. Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system. Biomed Microdevices 16, 375–385 (2014). https://doi.org/10.1007/s10544-014-9841-9

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