A microdevice for rapid, monoplex and colorimetric detection of foodborne pathogens using a centrifugal microfluidic platform
A microdevice for rapid, monoplex and colorimetric detection of foodborne pathogens using a centrifugal microfluidic platform
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DOI：
10.1016/j.bios.2017.08.060
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发表时间：
2018-02-15
影响因子：
12.6
通讯作者：
Kwai Lin Thong
中科院分区：
文献类型：
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作者：
Sayad, Abkar;Ibrahim, Fatimah;Kwai Lin Thong
食源性疾病的爆发已成为全球性的健康问题;因此，已经进行了许多改进和发展，以减少食品污染的风险。我们开发了一种离心式微流控自动无线终点检测系统，结合环介导等温扩增（LAMP）技术用于单一病原体检测。在微流控光盘（CD）上设计了6个相同的集，对3种不同的食源性病原体进行30次遗传分析。LAMP引物/试剂和DNA样品溶液的连续加载、混合和等分是使用优化的方波微通道、计量室和每分钟转数（RPM）控制来完成的。我们测试了24株致病菌（大肠杆菌，沙门氏菌和霍乱弧菌），每种细菌8株，并在微流控CD上进行DNA扩增60 min。然后，使用钙黄绿素比色法检测LAMP反应的扩增子，并通过开发的电子系统与蓝牙无线技术连接进行进一步分析，将结果传输到智能手机。该系统显示的检测限（LOD）为3 × 10（-5）ng μ L-1 DNA通过分析颜色变化时，与鸡肉与三种致病菌加标测试。由于整个过程是以全自动化的方式进行的，并且易于使用，因此我们的微型设备适用于非常简单的床旁（POC）测试，即使在资源有限的贫困环境中也能提供经济实惠和可访问性。
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 colt, Salmonella spp and Vibrio cholerae), with 8 strains of each bacterium, and performed DNA amplification on the microfluidic CD for 60 min. 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 x 10(-5) ng mu L-1 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.