title: Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system.
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.
authors.affiliation: Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA, melanie.hoehl@de.bosch.com.
authors.forename: Melanie M
authors.initials: MM
authors.lastname: Hoehl
authors.forename: Michael
authors.initials: M
authors.lastname: Weißert
authors.forename: Arne
authors.initials: A
authors.lastname: Dannenberg
authors.forename: Thomas
authors.initials: T
authors.lastname: Nesch
authors.forename: Nils
authors.initials: N
authors.lastname: Paust
authors.forename: Felix
authors.initials: F
authors.lastname: von Stetten
authors.forename: Roland
authors.initials: R
authors.lastname: Zengerle
authors.forename: Alexander H
authors.initials: AH
authors.lastname: Slocum
authors.forename: Juergen
authors.initials: J
authors.lastname: Steigert
mesh_terms: D001331:Automation; D002498:Centrifugation / Q000295:instrumentation*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D004209:Disposable Equipment; D011211:Electric Power Supplies; D005504:Food Analysis; D006361:Heating / Q000191:economics* / Q000295:instrumentation*; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D054323:Shiga-Toxigenic Escherichia coli / Q000166:cytology; D018511:Systems Integration
chemical_list: D004247:DNA