Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system
Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system
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DOI：
10.1007/s10544-014-9841-9
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发表时间：
2014-06-01
影响因子：
2.8
通讯作者：
Steigert, Juergen
中科院分区：
文献类型：
--
作者：
Hoehl, Melanie M.;Weissert, Michael;Steigert, Juergen
本文介绍了一种一次性电池驱动的加热系统，用于离心驱动的DNA纯化平台（LabTube）内的环介导等温DNA扩增（LAMP）。我们展示了基于LabTube的全自动DNA纯化，在实验室离心机中，在水，牛奶和苹果汁中低至100个细胞当量的产维罗毒素大肠杆菌（VTEC），然后进行集成和自动化的LAMP扩增，减少人手-加热系统由两个并联的SMD厚膜电阻和一个NTC作为加热和温度传感元件组成。它们由3V电池驱动，并由微控制器控制。将LAMP试剂储存在洗脱室中，并在将洗脱液吹扫到洗脱室中后立即开始扩增。LabTube包括一个基于微型离心机的加热系统，演示了在实验室离心机内进行无污染和自动化的样品到答案的核酸检测。加热系统可以很容易地在一个LabTube内并行化，并且可以部署用于各种加热和电气应用。
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.