Real-time gene analysis based on a portable electrochemical microfluidic system
Real-time gene analysis based on a portable electrochemical microfluidic system
复制标题
基于便携式电化学微流控系统的实时基因分析
DOI：
10.1016/j.elecom.2020.106665
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发表时间：
2020-02-01
影响因子：
5.4
通讯作者：
Li, Bingling
中科院分区：
文献类型：
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作者：
Liu, Yichen;Lu, Baiyang;Li, Bingling
等温扩增，例如环介导的核酸等温扩增 (LAMP)，是一种非常有前途的技术，可以彻底改变便携式或床旁基因检测。然而，这些等温反应常用的读数仅限于少数选项，例如实时荧光和比色纸条，它们分别在进一步积分和定量方面遇到实际困难。为了丰富读出库并提供更多适合各种检测要求的选项，我们报告了一种基于在封闭式便携式 PGE-LAMP 电化学芯片上执行等温核酸扩增反应的即用型基因测试方法。该方法以粪便污染物拟杆菌的HF183基因为模型靶点，实现了基因组信息到电化学信号的终点实时转导，具有超高的灵敏度、特异性和良好的信噪比，检测限低至80个拷贝。因此，这种方法对于即时检测系统的开发和加速 LAMP 的实际应用非常有前景。
Isothermal amplification, such as loop-mediated nucleic acid isothermal amplification (LAMP), is a highly promising technique that could revolutionize portable or point-of-care gene detection. However, the commonly used readouts of these isothermal reactions have been limited to a small number of options such as real-time fluorescence and colorimetric paper strips, which suffer from practical difficulties in further integration and quantitation, respectively. To enrich the readout library and provide more options suitable for a variety of detection requirements, we report a ready-to-use gene testing method based on the execution of isothermal nucleic acid amplification reactions on closed and portable PGE-LAMP electrochemical chips. Taking the HF183 gene of the fecal pollutant Bacteroides as a model target, this method allows both end-point and real-time transduction from genomic information to electrochemical signals with ultra-high sensitivity, specificity and a good signal-to-noise ratio, and with a detection limit as low as 80 copies. This approach is therefore highly promising for the development of point-of-care detection systems and accelerating the practical application of LAMP.