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一种基于免疫磁分离、酶催化和距离指示的用于可视化检测食源性病原体的微流控免疫传感器。

A microfluidic immunosensor for visual detection of foodborne bacteria using immunomagnetic separation, enzymatic catalysis and distance indication.

机构信息

Key Laboratory of Agricultural Information Acquisition Technology, Ministry of Agriculture and Rural Affairs, China Agricultural University, Beijing, 100083, China.

Key Laboratory of Modern Precision Agriculture System Integration Research, Ministry of Education, China Agricultural University, Beijing, 100083, China.

出版信息

Mikrochim Acta. 2019 Nov 9;186(12):757. doi: 10.1007/s00604-019-3883-x.

DOI:10.1007/s00604-019-3883-x
PMID:31707541
Abstract

A disposable visual microfluidic immunosensor is described for the determination of foodborne pathogens using immunomagnetic separation, enzymatic catalysis and distance indication. Specifically, a sensor was designed to detect Salmonella typhimurium as a model pathogen. Magnetic nanoparticles (MNPs) were modified with the anti-Salmonella monoclonal antibodies and then used to enrich S. typhimurium from the sample. This is followed by conjugation to polystyrene microspheres modified with anti-Salmonella polyclonal antibodies and catalase to form the MNP-bacteria-polystyrene-catalase sandwich. The catalase on the complexes catalyzes the decomposition of hydrogen peroxide to produce oxygen after passing a micromixer. The generated oxygen gas increases the pressure in the chip and pushes the indicating red dye solution to travel along the channel towards the unsealed outlet. The travel distance of the red dye can be visually read and related to the amount of S. typhimurium using the calibration scale. The sensor can detect as low as 150 CFU·mL within 2 h. Graphical abstractSchematic representation of the distance-based microfluidic immunosensor for visual detection of foodborne bacteria using immunomagnetic nanoparticles for bacteria separation, catalase for decomposition of hydrogen peroxide to form oxygen which causes a pressure increase, and red dyed particles movement for distance indication.

摘要

一种一次性可视化微流控免疫传感器,用于通过免疫磁分离、酶催化和距离指示来检测食源性病原体。具体而言,设计了一种传感器来检测鼠伤寒沙门氏菌作为模型病原体。磁性纳米颗粒 (MNP) 用抗沙门氏菌单克隆抗体修饰,然后用于从样品中富集鼠伤寒沙门氏菌。接下来,与聚苯乙烯微球共轭,该微球用抗沙门氏菌多克隆抗体和过氧化物酶修饰,形成 MNP-细菌-聚苯乙烯-过氧化物酶三明治结构。复合物上的过氧化物酶在通过微混合器后催化过氧化氢分解产生氧气。产生的氧气增加了芯片中的压力,并推动指示红色染料溶液沿着通道朝向未密封的出口移动。红色染料的行进距离可以通过目视读取,并使用校准刻度与鼠伤寒沙门氏菌的数量相关联。该传感器可以在 2 小时内检测到低至 150 CFU·mL 的细菌。

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