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A lab-on-chip device for the sample-in-result-out detection of viable Salmonella using loop-mediated isothermal amplification and real-time turbidity monitoring.
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-05-19 , DOI: 10.1039/d0lc00290a Siyuan Wang 1 , Ning Liu , Lingyan Zheng , Gaozhe Cai , Jianhan Lin
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-05-19 , DOI: 10.1039/d0lc00290a Siyuan Wang 1 , Ning Liu , Lingyan Zheng , Gaozhe Cai , Jianhan Lin
Affiliation
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Rapid screening of foodborne pathogens is key to prevent food poisoning. In this study, a lab-on-chip device was developed for rapid, automatic and sensitive detection of viable Salmonella typhimurium using loop-mediated isothermal amplification (LAMP) and smartphone real-time turbidity monitoring. First, magnetic nanoparticles (MNPs) coated with anti-Salmonella capture antibodies in propidium monoazide (PMA) were fully mixed with bacterial samples using two active magnetic stirring mixers at reverse rotating directions, and incubated in the serpentine channel with 470 nm blue light exposure, allowing specific formation of magnetic bacteria and sufficient PMA pretreatment of the DNA of dead bacteria. Then, the PMA-treated magnetic bacteria were separated in the separation chamber using the magnetic field and their genomic DNA templates were extracted using lysis buffer at 70 °C. Finally, the viable bacteria's DNA was amplified using LAMP in the detection chamber preloaded with the lyophilized LAMP reagents at 67.5 °C after blocking with paraffin oil to avoid aerosol cross contamination. Finally, the turbidity of the LAMP reaction system was monitored in a real-time manner for the quantitative detection of viable bacteria. The experimental results demonstrated that this device was able to automatically detect viable Salmonella as low as 14 CFU mL−1 in spiked chicken meat supernatants within 1.5 h. This device is very promising to provide a sample-in-result-out solution for the in-field detection of Salmonella and could be easily extended for other foodborne pathogens.
中文翻译：
一种利用回路介导的等温扩增和实时浊度监测功能进行活检沙门氏菌样品进样检测的芯片实验室设备。
快速筛选食源性病原体是预防食物中毒的关键。在这项研究中，开发了一种芯片实验室设备，用于使用回路介导的等温扩增（LAMP）和智能手机实时浊度监测来快速，自动和灵敏地检测鼠伤寒沙门氏菌。首先，涂有抗沙门氏菌的磁性纳米颗粒（MNP）使用两个主动磁力搅拌混合器以反向旋转将单叠氮化丙锭（PMA）中的捕获抗体与细菌样品充分混合，并在蛇形通道中暴露于470 nm的蓝光下进行孵育，从而特异性形成磁性细菌并对其进行足够的PMA预处理死亡细菌的DNA。然后，使用磁场在分离室中分离经PMA处理的磁性细菌，并使用70°C的裂解缓冲液提取其基因组DNA模板。最后，在用石蜡油封闭以避免气溶胶交叉污染后，在预先装有冻干的LAMP试剂的检测室中，用LAMP在67.5°C下扩增LAMP，扩增活细菌的DNA。最后，实时监测LAMP反应系统的浊度，以定量检测活菌。实验结果表明，该设备能够自动检测可行在1.5小时内，加标鸡肉上清液中的沙门氏菌低至14 CFU mL -1。该设备非常有希望为野外检测沙门氏菌提供一个结果输入解决方案，并且可以很容易地扩展到其他食源性病原体。
更新日期：2020-06-30
中文翻译：
一种利用回路介导的等温扩增和实时浊度监测功能进行活检沙门氏菌样品进样检测的芯片实验室设备。
快速筛选食源性病原体是预防食物中毒的关键。在这项研究中，开发了一种芯片实验室设备，用于使用回路介导的等温扩增（LAMP）和智能手机实时浊度监测来快速，自动和灵敏地检测鼠伤寒沙门氏菌。首先，涂有抗沙门氏菌的磁性纳米颗粒（MNP）使用两个主动磁力搅拌混合器以反向旋转将单叠氮化丙锭（PMA）中的捕获抗体与细菌样品充分混合，并在蛇形通道中暴露于470 nm的蓝光下进行孵育，从而特异性形成磁性细菌并对其进行足够的PMA预处理死亡细菌的DNA。然后，使用磁场在分离室中分离经PMA处理的磁性细菌，并使用70°C的裂解缓冲液提取其基因组DNA模板。最后，在用石蜡油封闭以避免气溶胶交叉污染后，在预先装有冻干的LAMP试剂的检测室中，用LAMP在67.5°C下扩增LAMP，扩增活细菌的DNA。最后，实时监测LAMP反应系统的浊度，以定量检测活菌。实验结果表明，该设备能够自动检测可行在1.5小时内，加标鸡肉上清液中的沙门氏菌低至14 CFU mL -1。该设备非常有希望为野外检测沙门氏菌提供一个结果输入解决方案，并且可以很容易地扩展到其他食源性病原体。
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