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

被引:14
作者
Hoehl, Melanie M. [1 ,2 ,3 ]
Weissert, Michael [3 ,4 ]
Dannenberg, Arne [3 ]
Nesch, Thomas [5 ]
Paust, Nils [6 ]
von Stetten, Felix [6 ]
Zengerle, Roland [6 ]
Slocum, Alexander H. [1 ,2 ]
Steigert, Juergen [3 ]
机构
[1] MIT, Harvard MIT Div Hlth Sci & Technol, Cambridge, MA 02139 USA
[2] MIT, Dept Mech Engn, Cambridge, MA 02139 USA
[3] Robert Bosch GmbH, Adv Res & Engn, Gerlingen, Germany
[4] Univ Stuttgart, D-70174 Stuttgart, Germany
[5] Nesch Engn UG Haftungsbeschrankt, Haigerloch, Germany
[6] HSG IMIT, Freiburg, Germany
关键词
Automated DNA extraction; DNA amplification; LAMP; Heater; Battery; MEDIATED ISOTHERMAL AMPLIFICATION; VIABLE ESCHERICHIA-COLI; PATHOGEN DETECTION; RAPID DETECTION; NUCLEIC-ACIDS; DRINKING-WATER; EXTRACTION; BIOSENSORS; PCR; DEVICES;
D O I
10.1007/s10544-014-9841-9
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
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.
引用
收藏
页码:375 / 385
页数:11
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