Yu Mengtao, Huang Pei, Li Yuanguo, Song Yumeng, Liu Xingqi, Feng Na, Jin Hongli, Bai Yujie, Zhang Haili, Li Yuanyuan, Xia Xianzhu, Gao Yuwei, Wang Hualei
State Key Laboratory for Zoonotic Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun, China.
Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, China.
Front Microbiol. 2022 Jul 13;13:932698. doi: 10.3389/fmicb.2022.932698. eCollection 2022.
SARS-CoV-2 is a novel coronavirus that has caused a global pandemic. To date, 504,907,616 people have been infected and developed coronavirus disease 2019 (COVID-19). A rapid and simple diagnostic method is needed to control this pandemic. In this study, a visual nucleic acid detection method combining reverse transcription loop-mediated isothermal amplification and a vertical flow visualization strip (RT-LAMP-VF) was successfully established and could detect 20 copies/μl of SARS-CoV-2 RNA transcript within 50 min at 61°C. This assay had no cross-reactivity with a variety of coronaviruses, including human coronavirus OC43, 229E, HKU1, NL63, severe acute respiratory syndrome-related coronavirus (SARSr-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and bat coronavirus HKU4, exhibiting very high levels of diagnostic sensitivity and specificity. Most strikingly, this method can be used for detecting multiple SARS-CoV-2 variants, including the Wuhan-Hu-1 strain, Delta, and Omicron variants. Compared with the RT-qPCR method recommended by the World Health Organization (WHO), RT-LAMP-VF does not require special equipment and is easy to perform. As a result, it is more suitable for rapid screening of suspected SARS-CoV-2 samples in the field and local laboratories.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是一种新型冠状病毒，已引发全球大流行。截至目前，已有504,907,616人感染并患上2019冠状病毒病（COVID-19）。需要一种快速简便的诊断方法来控制这一疫情。在本研究中，成功建立了一种将逆转录环介导等温扩增与垂直流可视化试纸条相结合的可视化核酸检测方法（RT-LAMP-VF），该方法在61°C下50分钟内可检测到20拷贝/μl的SARS-CoV-2 RNA转录本。该检测方法与多种冠状病毒无交叉反应，包括人冠状病毒OC43、229E、HKU1、NL63、严重急性呼吸综合征相关冠状病毒（SARSr-CoV）、中东呼吸综合征冠状病毒（MERS-CoV）和蝙蝠冠状病毒HKU4，具有很高的诊断敏感性和特异性。最引人注目的是，该方法可用于检测多种SARS-CoV-2变异株，包括武汉-胡-1株、德尔塔变异株和奥密克戎变异株。与世界卫生组织（WHO）推荐的RT-qPCR方法相比，RT-LAMP-VF不需要特殊设备，操作简便。因此，它更适合在现场和当地实验室对疑似SARS-CoV-2样本进行快速筛查。
