DeFina Samuel M, Wang Jianhui, Yang Lei, Zhou Han, Adams Jennifer, Cushing William, Tuohy Beth, Hui Pei, Liu Chen, Pham Kien
Department of Pathology, Yale School of Medicine, Yale University, New Haven, CT, USA.
Department of Laboratory Medicine, Yale School of Medicine, Yale University, New Haven, CT, USA.
Sci Rep. 2022 Apr 6;12(1):5729. doi: 10.1038/s41598-022-09718-4.
The Coronavirus disease 2019 (COVID-19) pandemic-caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)- has posed a global threat and presented with it a multitude of economic and public-health challenges. Establishing a reliable means of readily available, rapid diagnostic testing is of paramount importance in halting the spread of COVID-19, as governments continue to ease lockdown restrictions. The current standard for laboratory testing utilizes reverse transcription quantitative polymerase chain reaction (RT-qPCR); however, this method presents clear limitations in requiring a longer run-time as well as reduced on-site testing capability. Therefore, we investigated the feasibility of a reverse transcription looped-mediated isothermal amplification (RT-LAMP)-based model of rapid COVID-19 diagnostic testing which allows for less invasive sample collection, named SaliVISION. This novel, two-step, RT-LAMP assay utilizes a customized multiplex primer set specifically targeting SARS-CoV-2 and a visual report system that is ready to interpret within 40 min from the start of sample processing and does not require a BSL-2 level testing environment or special laboratory equipment. When compared to the SalivaDirect and Thermo Fisher Scientific TaqPath RT-qPCR testing platforms, the respective sensitivities of the SaliVISION assay are 94.29% and 98.28% while assay specificity was 100% when compared to either testing platform. Our data illustrate a robust, rapid diagnostic assay in our novel RT-LAMP test design, with potential for greater testing throughput than is currently available through laboratory testing and increased on-site testing capability.
2019年冠状病毒病（COVID-19）由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起，已构成全球威胁，并带来了众多经济和公共卫生挑战。随着各国政府继续放宽封锁限制，建立一种可靠的、随时可用的快速诊断检测方法对于遏制COVID-19的传播至关重要。目前实验室检测的标准方法是逆转录定量聚合酶链反应（RT-qPCR）；然而，这种方法存在明显局限性，需要较长的运行时间，且现场检测能力有限。因此，我们研究了一种基于逆转录环介导等温扩增（RT-LAMP）的快速COVID-19诊断检测模型的可行性，该模型允许采用侵入性较小的样本采集方式，名为SaliVISION。这种新颖的两步RT-LAMP检测方法使用了专门针对SARS-CoV-2的定制多重引物组和一个视觉报告系统，该系统在样本处理开始后40分钟内即可解读结果，且不需要生物安全二级（BSL-2）检测环境或特殊实验室设备。与SalivaDirect和赛默飞世尔科技的TaqPath RT-qPCR检测平台相比，SaliVISION检测方法的灵敏度分别为94.29%和98.28%，与任一检测平台相比，检测特异性均为100%。我们的数据表明，我们新颖的RT-LAMP检测设计是一种强大的快速诊断检测方法，具有比目前实验室检测更高的检测通量潜力和更强的现场检测能力。
