A fully automated centrifugal microfluidic system for sample-to-answer viral nucleic acid testing
A fully automated centrifugal microfluidic system for sample-to-answer viral nucleic acid testing
复制标题
用于从样本到答案的病毒核酸检测的全自动离心微流体系统
DOI：
10.1007/s11426-020-9800-6
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发表时间：
2020-07-27
影响因子：
9.6
通讯作者：
Sun, Jiashu
中科院分区：
文献类型：
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作者：
Tian, Fei;Liu, Chao;Sun, Jiashu
病毒引起的传染病的爆发构成了全球公共卫生挑战。核酸扩增检测 (NAAT) 能够及早发现大流行病毒，并在防止进一步传播方面发挥重要作用。然而，对熟练的操作人员、昂贵的仪器和生物安全实验室的要求阻碍了NAAT用于疑似患者的筛查和诊断。在此，我们报告了一种全自动离心微流体系统的开发，该系统具有样本输入-应答输出功能，可用于敏感、特异和快速的病毒核酸检测。核酸的释放和随后的逆转录环介导的等温扩增（RT-LAMP）被集成到微流体盘的反应单元中。试剂注入、旋转流体驱动、加热控温、荧光信号检测等整个处理步骤均由定制仪器自动完成。我们使用掺有严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 装甲 RNA 颗粒的口咽拭子样本验证了离心微流体系统。装甲 RNA 颗粒的估计检测限为每个反应 2 个拷贝，吞吐量为每个盘 21 个反应，检测样本到给出结果的时间约为 70 分钟。这种封闭的自动化微流体系统有效地避免了气溶胶的病毒污染，并且可以很容易地适用于诊断实验室外的病毒检测。
The outbreak of virus-induced infectious diseases poses a global public-health challenge. Nucleic acid amplification testing (NAAT) enables early detection of pandemic viruses and plays a vital role in preventing onward transmission. However, the requirement of skilled operators, expensive instrumentation, and biosafety laboratories has hindered the use of NAAT for screening and diagnosis of suspected patients. Here we report development of a fully automated centrifugal microfluidic system with sample-in-answer-out capability for sensitive, specific, and rapid viral nucleic acid testing. The release of nucleic acids and the subsequent reverse transcription loop-mediated isothermal amplification (RT-LAMP) were integrated into the reaction units of a microfluidic disc. The whole processing steps such as injection of reagents, fluid actuation by rotation, heating and temperature control, and detection of fluorescence signals were carried out automatically by a customized instrument. We validate the centrifugal microfluidic system using oropharyngeal swab samples spiked with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) armored RNA particles. The estimated limit of detection for armored RNA particles is 2 copies per reaction, the throughput is 21 reactions per disc, and the assay sample-to-answer time is approximately 70 min. This enclosed and automated microfluidic system efficiently avoids viral contamination of aerosol, and can be readily adapted for virus detection outside the diagnostic laboratory.