Van Ngoc Huynh, Quyen Than Linh, Vinayaka Aaydha Chidambara, Bang Dang Duong, Wolff Anders
BioLabChip Group, Department of Biotechnology and Biomedicine, Technical University of Denmark (DTU-Bioengineering), Lyngby, Denmark.
Laboratory of Applied Micro and Nanotechnology (LAMINATE), Department of Biotechnology and Biomedicine, Technical University of Denmark (DTU-Bioengineering), Lyngby, Denmark.
Front Bioeng Biotechnol. 2022 Aug 4;10:917573. doi: 10.3389/fbioe.2022.917573. eCollection 2022.
The COVID-19 pandemic emphasized the importance of rapid, portable, and on-site testing technologies necessary for resource-limited settings for effective testing and screening to reduce spreading of the infection. Realizing this, we developed a fluorescence-based point-of-care (fPOC) detection system with real-time reverse transcriptase loop-mediated isothermal amplification for rapid and quantitative detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. The system is built based on the Arduino platform compatible with commercially available open-source hardware-software and off-the-shelf electronic components. The fPOC system comprises of three main components: 1) an instrument with integrated heaters, 2) optical detection components, and 3) an injection-molded polymeric cartridge. The system was tested and experimentally proved to be able to use for fast detection of the SARS-CoV-2 virus in real-time in less than 30 min. Preliminary results of testing the performance of the fPOC revealed that the fPOC could detect the SARS-CoV-2 virus at a limit of detection (LOD) at two to three copies/microliter (15.36 copies/reaction), which was comparable to reactions run on a standard commercial thermocycler. The performance of the fPOC was evaluated with 12 SARS-CoV-2 clinical throat swab samples that included seven positive and five negative samples, as confirmed by reverse transcription-polymerase chain reaction. The fPOC showed 100% agreement with the commercial thermocycler. This simple design of the fPOC system demonstrates the potential to greatly enhance the practical applicability to develop a totally integrated point-of-care system for rapid on-site screening of the SARS-CoV-2 virus in the management of the pandemic.
新冠疫情凸显了快速、便携和现场检测技术对于资源有限地区进行有效检测和筛查以减少感染传播的重要性。认识到这一点后,我们开发了一种基于荧光的即时检测(fPOC)系统,该系统采用实时逆转录环介导等温扩增技术,用于快速定量检测严重急性呼吸综合征冠状病毒2(SARS-CoV-2)病毒。该系统基于与市售开源硬件软件和现成电子元件兼容的Arduino平台构建。fPOC系统由三个主要组件组成:1)带有集成加热器的仪器,2)光学检测组件,3)注塑聚合物盒。该系统经过测试,实验证明能够在不到30分钟的时间内实时快速检测SARS-CoV-2病毒。对fPOC性能进行测试的初步结果显示,fPOC能够以每微升两到三个拷贝(15.36个拷贝/反应)的检测限检测SARS-CoV-2病毒,这与在标准商用热循环仪上进行的反应相当。通过逆转录聚合酶链反应确认,使用12份SARS-CoV-2临床咽拭子样本(包括7份阳性和5份阴性样本)对fPOC的性能进行了评估。fPOC与商用热循环仪的检测结果完全一致。fPOC系统这种简单的设计表明,在疫情管理中开发一个完全集成的即时检测系统用于快速现场筛查SARS-CoV-2病毒具有极大提高实际适用性的潜力。