Viscometric real-time monitoring of the rolling circle amplification with a micromachined cantilever
Viscometric real-time monitoring of the rolling circle amplification with a micromachined cantilever
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DOI：
10.1016/j.snb.2016.08.105
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发表时间：
2017-02-01
影响因子：
8.4
通讯作者：
Dual, Jurg
中科院分区：
文献类型：
--
作者：
Rust, Philipp;Cereghetti, Damiano;Dual, Jurg
DNA扩增反应通常用于芯片实验室装置上的核酸分析。考虑到例如用于急救站中的病原体检测的护理点测试，这样的分析必须快速且简单地执行。作为基于荧光的测量的替代方法，本文提出的方法基于检测测定的流变学性质的变化。对于第一次，在芯片上的DNA扩增与真实的时间粘度检测显示，与第一个结果已经可在不到20分钟后。在聚合过程中，采用滚环扩增（RCA）的样品的粘度的变化检测使用微机械振动悬臂。选择RCA，因为它是等温的并且产生非常长的股线，导致流体的流变性质的高度变化。随着反应的进行，实时观察到悬臂的阻尼和保持其振幅恒定所需的激励功率的增加。评价这两个参数的斜率，例如在反应开始后27分钟，可以完全区分发生聚合的样品和没有反应的样品。所描述的方法在没有传感器表面的明确官能化或反应组分的标记的情况下起作用。(C)2016爱思唯尔B.V.保留所有权利。
DNA amplification reactions are often used for nucleic acid analysis on lab-on-chip devices. In view of point of care testing e.g. for pathogen detection in emergency stations, such an analysis must be fast and simple to be performed. As an alternative to fluorescence based measurements the method presented here is based on detecting the change in the rheological properties of the assay. For the first time, on chip DNA amplification with real time viscometric detection is shown, with first results already being available after less than 20 min. The change of the sample's viscosity during the polymerization employing the rolling circle amplification (RCA) is detected using a micromachined vibrating cantilever. The RCA was chosen, as it is isothermal and produces very long strands resulting in a high change of the fluid's rheological properties. As the reaction proceeds, an increase of the cantilever's damping and required excitation power to keep its amplitude constant is observed in real-time. Evaluating the slope of these two parameters e.g. 27 min after starting the reaction, samples where polymerization takes place and samples without reaction can perfectly be distinguished. The described approach works without explicit functionalization of the sensor surface or labeling of reaction components. (C) 2016 Elsevier B.V. All rights reserved.