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聚多巴胺聚集:一种用于无电源读出环介导等温扩增的新策略,该策略集成于用于多重病原体检测的纸质装置中。

Polydopamine aggregation: A novel strategy for power-free readout of loop-mediated isothermal amplification integrated into a paper device for multiplex pathogens detection.

作者信息

Nguyen Hanh An, Lee Nae Yoon

机构信息

Department of BioNano Technology, Gachon University, 1342 Seongnam-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do, 13120, South Korea.

Department of BioNano Technology, Gachon University, 1342 Seongnam-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do, 13120, South Korea.

出版信息

Biosens Bioelectron. 2021 Oct 1;189:113353. doi: 10.1016/j.bios.2021.113353. Epub 2021 May 18.

DOI:10.1016/j.bios.2021.113353
PMID:34049080
Abstract

Loop-mediated isothermal amplification (LAMP) has been widely used for detecting pathogens. However, power-free and clear visualization of results still remain challenging. In this study, we developed a paper device integrated with power-free DNA detection strategy realized by polydopamine aggregation. In the presence of DNA amplicons, the polymerization of dopamine into aggregated polydopamine was hindered, while in the absence of DNA amplicons, polydopamine aggregation is facilitated. The porosity of the paper enabled the capillary flow of dispersed polydopamine for positive sample, while aggregated polydopamine remained at the bottom of the paper strip due to large size of the aggregates for negative sample. Based on this mechanism, we fabricated a slidable paper device integrating LAMP with dopamine polymerization for the naked-eye detection, operated in a seamless manner. Moreover, the introduced paper device was successfully used to detect DNA extracted from Escherichia coli O157:H7 and SARS-CoV-2 within 25 min, as well as Enterococcus faecium within 35 min. The detection limits of both Escherichia coli O157:H7 and SARS-CoV-2 were 10 ng/μL. The introduced paper device can be used as a simple and sensitive tool for detecting multiple infectious pathogens, making it an ideal tool particularly for resource-limited environment.

摘要

环介导等温扩增技术(LAMP)已被广泛用于病原体检测。然而,实现无电源且结果清晰可视化仍具有挑战性。在本研究中,我们开发了一种集成了通过聚多巴胺聚集实现的无电源DNA检测策略的纸质装置。在存在DNA扩增子的情况下,多巴胺聚合成聚集的聚多巴胺的过程受到阻碍,而在不存在DNA扩增子的情况下,聚多巴胺聚集则得到促进。纸张的孔隙率使得分散的聚多巴胺能够通过毛细作用流动用于阳性样本,而由于聚集物尺寸较大,聚集的聚多巴胺则留在纸条底部用于阴性样本。基于这一机制,我们制造了一种将LAMP与多巴胺聚合相结合的可滑动纸质装置,用于肉眼检测,操作无缝衔接。此外,所引入的纸质装置成功用于在25分钟内检测从大肠杆菌O157:H7和严重急性呼吸综合征冠状病毒2(SARS-CoV-2)中提取的DNA,以及在35分钟内检测粪肠球菌。大肠杆菌O157:H7和SARS-CoV-2的检测限均为10 ng/μL。所引入的纸质装置可作为一种简单且灵敏的工具用于检测多种传染性病原体,使其成为特别适用于资源有限环境的理想工具。

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