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一种使用离心微流控平台快速、单plex 和比色检测食源性病原体的微器件。

A microdevice for rapid, monoplex and colorimetric detection of foodborne pathogens using a centrifugal microfluidic platform.

机构信息

Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, Kuala Lumpur, Malaysia; Centre for Innovation in Medical Engineering (CIME), Faculty of Engineering, University of Malaya, Kuala Lumpur, Malaysia.

Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, Kuala Lumpur, Malaysia; Centre for Innovation in Medical Engineering (CIME), Faculty of Engineering, University of Malaya, Kuala Lumpur, Malaysia; Department of Biomedical Engineering, Inje University, Gimhae 621-749, South Korea.

出版信息

Biosens Bioelectron. 2018 Feb 15;100:96-104. doi: 10.1016/j.bios.2017.08.060. Epub 2017 Aug 31.

Abstract

Outbreaks of foodborne diseases have become a global health concern; hence, many improvements and developments have been made to reduce the risk of food contamination. We developed a centrifugal microfluidic automatic wireless endpoint detection system integrated with loop mediated isothermal amplification (LAMP) for monoplex pathogen detection. Six identical sets were designed on the microfluidic compact disc (CD) to perform 30 genetic analyses of three different species of foodborne pathogens. The consecutive loading, mixing, and aliquoting of the LAMP primers/reagents and DNA sample solutions were accomplished using an optimized square-wave microchannel, metering chambers and revulsion per minute (RPM) control. We tested 24 strains of pathogenic bacteria (Escherichia coli, Salmonella spp and Vibrio cholerae), with 8 strains of each bacterium, and performed DNA amplification on the microfluidic CD for 60min. Then, the amplicons of the LAMP reaction were detected using the calcein colorimetric method and further analysed via the developed electronic system interfaced with Bluetooth wireless technology to transmit the results to a smartphone. The system showed a limit of detection (LOD) of 3 × 10ngμL DNA by analysing the colour change when tested with chicken meat spiked with the three pathogenic bacteria. Since the entire process was performed in a fully automated way and was easy to use, our microdevice is suitable for point-of-care (POC) testing with high simplicity, providing affordability and accessibility even to poor, resource-limited settings.

摘要

食源性疾病的爆发已成为全球关注的健康问题;因此,为了降低食品污染的风险,已经进行了许多改进和发展。我们开发了一种离心微流自动无线终点检测系统,该系统与环介导等温扩增(LAMP)集成在一起,用于单重病原体检测。在微流控光盘(CD)上设计了六套相同的装置,用于对三种不同食源性病原体进行 30 次基因分析。使用优化的方波微通道、计量室和每分钟反转数(RPM)控制来完成 LAMP 引物/试剂和 DNA 样品溶液的连续加载、混合和分配。我们测试了 24 株病原菌(大肠杆菌、沙门氏菌和霍乱弧菌),每种细菌有 8 株,并在微流控 CD 上进行了 60min 的 DNA 扩增。然后,使用钙黄绿素比色法检测 LAMP 反应的扩增产物,并通过与蓝牙无线技术接口的开发电子系统进行进一步分析,将结果传输到智能手机上。该系统通过分析用三种病原菌污染的鸡肉时的颜色变化,显示出检测限(LOD)为 3×10ngμL DNA。由于整个过程是全自动进行的,并且易于使用,因此我们的微设备非常适合即时检测(POC)测试,具有高度的简单性,即使在贫穷、资源有限的环境中也具有经济实惠性和可及性。

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