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一种集成的旋转微流控系统,用于即时诊断点的病原体,具有 DNA 提取、环介导等温扩增和基于侧流条的检测。

An integrated rotary microfluidic system with DNA extraction, loop-mediated isothermal amplification, and lateral flow strip based detection for point-of-care pathogen diagnostics.

机构信息

Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.

Department of Chemical Engineering, College of Engineering, Kyung Hee University, 1 Seochon-dong, Giheung-gu, Yongin-si, Gyeonggi-do 17140, Republic of Korea.

出版信息

Biosens Bioelectron. 2017 May 15;91:334-340. doi: 10.1016/j.bios.2016.11.063. Epub 2016 Dec 14.

Abstract

Point-of-care (POC) molecular diagnostics plays a pivotal role for the prevention and treatment of infectious diseases. In spite of recent advancement in microfluidic based POC devices, there are still rooms for development to realize rapid, automatic and cost-effective sample-to-result genetic analysis. In this study, we propose an integrated rotary microfluidic system that is capable of performing glass microbead based DNA extraction, loop mediated isothermal amplification (LAMP), and colorimetric lateral flow strip based detection in a sequential manner with an optimized microfluidic design and a rotational speed control. Rotation direction-dependent coriolis force and siphon valving structures enable us to perform the fluidic control and metering, and the use of the lateral flow strip as a detection method renders all the analytical processes for nucleic acid test simplified and integrated without the need of expensive instruments or human intervention. As a proof of concept for point-of-care DNA diagnostics, we identified the food-borne bacterial pathogen which was contaminated in water or milk. Not only monoplex Salmonella Typhimurium but also multiplex Salmonella Typhimurium and Vibrio parahaemolyticus were analysed on the integrated rotary genetic analysis microsystem with a limit of detection of 50 CFU in 80min. In addition, three multiple samples were simultaneously analysed on a single device. The sample-to-result capability of the proposed microdevice provides great usefulness in the fields of clinical diagnostics, food safety and environment monitoring.

摘要

即时检测(POC)分子诊断在传染病的预防和治疗中起着关键作用。尽管基于微流控的即时检测设备最近取得了进展,但仍有发展空间,以实现快速、自动和具有成本效益的样本到结果遗传分析。在本研究中,我们提出了一种集成旋转微流控系统,该系统能够以优化的微流控设计和转速控制,顺序执行基于玻璃微珠的 DNA 提取、环介导等温扩增(LAMP)和基于比色横向流动条的检测。旋转方向相关的科里奥利力和虹吸管阀结构使我们能够进行流体控制和计量,而使用横向流动条作为检测方法使核酸测试的所有分析过程简化和集成,而无需昂贵的仪器或人工干预。作为即时护理 DNA 诊断的概念验证,我们鉴定了污染在水或牛奶中的食源性病原体。在集成旋转基因分析微系统上,不仅可以分析单重伤寒沙门氏菌 Typhimurium,还可以分析多重伤寒沙门氏菌 Typhimurium 和副溶血性弧菌,检测限为 50CFU,耗时 80 分钟。此外,三个多个样本可以同时在单个设备上进行分析。该微器件的样本到结果能力在临床诊断、食品安全和环境监测等领域具有很大的实用性。

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