肉眼
环介导等温扩增
核酸
化学
材料科学
纳米技术
生物物理学
生物系统
色谱法
生物化学
DNA
生物
检出限
作者
Mei Fang,Yiru Wang,Tao Yang,Jing Zhang,Hanry Yu,Zisheng Luo,Bin Su,Xingyu Lin
出处
期刊:ACS Sensors
[American Chemical Society]
日期:2024-04-11
卷期号:9 (4): 2010-2019
被引量:1
标识
DOI:10.1021/acssensors.3c02807
摘要
Digital nucleic acid amplification enables the absolute quantification of single molecules. However, due to the ultrasmall reaction volume in the digital system (i.e., short light path), most digital systems are limited to fluorescence signals, while label-free and naked-eye readout remain challenging. In this work, we report a digital nucleic acid plate culture method for label-free, ultrasimple, and naked-eye nucleic acid analysis. As simple as the bacteria culture, the nanoconfined digital loop-mediated isothermal amplification was performed by using polyacrylamide (PAM) hydrogel as the amplification matrix. The nanoconfinement of PAM hydrogel with an ionic polymer chain can remarkably accelerate the amplification of target nucleic acids and the growth of inorganic byproducts, namely, magnesium pyrophosphate particles (MPPs). Compared to that in aqueous solutions, MPPs trapped in the hydrogel with enhanced light scattering characteristics are clearly visible to the naked eye, forming white "colony" spots that can be simply counted in a label-free and instrument-free manner. The MPPs can also be photographed by a smartphone and automatically counted by a machine-learning algorithm to realize the absolute quantification of antibiotic-resistant pathogens in diverse real samples.
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