Li Nan, Shen Minjie, Liu Jiajia, Zhang Li, Wang Huili, Xu Youchun, Cheng Jing
State Key Laboratory of Membrane Biology, Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing, 100084 China.
National Engineering Research Center for Beijing Biochip Technology, Beijing, 102206 China.
Microsyst Nanoeng. 2021 Nov 23;7:94. doi: 10.1038/s41378-021-00321-7. eCollection 2021.
Coronavirus disease 2019 (COVID-19) has emerged, rapidly spread and caused significant morbidity and mortality worldwide. There is an urgent public health need for rapid, sensitive, specific, and on-site diagnostic tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In this study, a fully integrated and portable analyzer was developed to detect SARS-CoV-2 from swab samples based on solid-phase nucleic acid extraction and reverse transcription loop-mediated isothermal amplification (RT-LAMP). The swab can be directly inserted into a cassette for multiplexed detection of respiratory pathogens without pre-preparation. The overall detection process, including swab rinsing, magnetic bead-based nucleic acid extraction, and 8-plex real-time RT-LAMP, can be automatically performed in the cassette within 80 min. The functionality of the cassette was validated by detecting the presence of a SARS-CoV-2 pseudovirus and three other respiratory pathogens, i.e., , , and . The limit of detection (LoD) for the SARS-CoV-2 pseudovirus was 2.5 copies/μL with both primer sets ( gene and gene), and the three bacterial species were successfully detected with an LoD of 2.5 colony-forming units (CFU)/μL in 800 μL of swab rinse. Thus, the analyzer developed in this study has the potential to rapidly detect SARS-CoV-2 and other respiratory pathogens on site in a "raw-sample-in and answer-out" manner.
2019年冠状病毒病（COVID-19）已出现，并在全球迅速传播，导致了严重的发病和死亡情况。对于严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染，迫切需要快速、灵敏、特异且可现场操作的诊断检测方法。在本研究中，开发了一种完全集成的便携式分析仪，基于固相核酸提取和逆转录环介导等温扩增（RT-LAMP）技术，用于从拭子样本中检测SARS-CoV-2。拭子可直接插入检测卡盒，无需预先准备即可对呼吸道病原体进行多重检测。整个检测过程，包括拭子冲洗、磁珠法核酸提取以及8重实时RT-LAMP，可在检测卡盒内自动完成，耗时80分钟。通过检测SARS-CoV-2假病毒以及其他三种呼吸道病原体（即 、 和 ）的存在，验证了检测卡盒的功能。两种引物组（ 基因和 基因）对SARS-CoV-2假病毒的检测限均为2.5拷贝/微升，在800微升的拭子冲洗液中，三种细菌均以2.5菌落形成单位（CFU）/微升的检测限成功检测到。因此，本研究开发的分析仪有潜力以“样本进、结果出”的方式现场快速检测SARS-CoV-2和其他呼吸道病原体。
