Center for Molecular Recognition and Biosensing, School of Life Sciences, Shanghai University, Shanghai, 200444, China.
CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, 215163, Jiangsu, China.
Anal Bioanal Chem. 2019 Sep;411(23):6039-6047. doi: 10.1007/s00216-019-01985-x. Epub 2019 Jul 15.
Circulating tumor DNA (ctDNA) is a tumor-derived fragmented DNA in the bloodstream that is not associated with cells. It has been greatly focused in the recent decade because of its potential clinical utility for liquid biopsies. Development of ctDNA analytical techniques with high sensitivity and cost-efficiency will undoubtedly promote the clinical spread of ctDNA testing. In this paper, we propose a novel flow cytometry-based ctDNA sensing strategy which combines enzyme-free amplification and magnetic separation. The target DNA is capable of triggering a hybridization chain reaction, producing a fluorescent long linear assembly of DNA, which can be further captured by magnetic beads to present fluorescent signals using flow cytometry. In comparison with some conventional methods, our strategy has the advantages of easy operation and cost-efficiency, and thereby shows a promising application in clinical diagnosis. Graphical abstract.
循环肿瘤 DNA(ctDNA)是血液中与细胞无关的肿瘤源性片段化 DNA。由于其在液体活检方面的潜在临床应用,近年来受到了极大的关注。开发具有高灵敏度和成本效益的 ctDNA 分析技术无疑将促进 ctDNA 检测的临床应用。在本文中,我们提出了一种基于流动 cytometry 的新型 ctDNA 传感策略,该策略结合了无酶扩增和磁分离。靶 DNA 能够触发杂交链式反应,产生荧光长线性 DNA 组装体,然后使用流动 cytometry 通过磁性珠进一步捕获,以呈现荧光信号。与一些传统方法相比,我们的策略具有操作简单和成本效益高的优点,因此在临床诊断中具有广阔的应用前景。