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

• Published in: Biomedical microdevices Vol. 16; no. 3; pp. 375 - 385
• Main Authors: Hoehl, Melanie M., Weißert, Michael, Dannenberg, Arne, Nesch, Thomas, Paust, Nils, von Stetten, Felix, Zengerle, Roland, Slocum, Alexander H., Steigert, Juergen
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.06.2014; Springer Nature B.V
• Subjects: Automation; Batteries; Biological and Medical Physics; Biomedical engineering; Biomedical Engineering and Bioengineering; Biomedical materials; Biophysics; Centrifugation - instrumentation; Disposable Equipment; DNA - genetics; DNA - isolation & purification; Electric Power Supplies; Engineering; Engineering Fluid Dynamics; Escherichia coli; Food Analysis; Heating - economics; Heating - instrumentation; Loops; Malus; Nanotechnology; Polymerase Chain Reaction - instrumentation; Shiga-Toxigenic Escherichia coli - cytology; Systems Integration; DNA amplification; LAMP; Automated DNA extraction; Heater; Battery
• ISSN: 1387-2176; 1572-8781
• DOI: 10.1007/s10544-014-9841-9

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.