Borah Slater Khushboo, Ahmad Muhammad, Poirier Aurore, Stott Ash, Siedler Bianca Sica, Brownsword Matthew, Mehat Jai, Urbaniec Joanna, Locker Nicolas, Zhao Yunlong, La Ragione Roberto, Silva S Ravi P, McFadden Johnjoe
School of Biosciences, Faculty of Health and Medical Sciences, University of Surrey, Guildford GU2 7XH, UK.
Advanced Technology Institute, University of Surrey, Guildford GU2 7XH, UK.
iScience. 2023 Aug 9;26(9):107570. doi: 10.1016/j.isci.2023.107570. eCollection 2023 Sep 15.
Rapid, reliable, sensitive, portable, and accurate diagnostics are required to control disease outbreaks such as COVID-19 that pose an immense burden on human health and the global economy. Here we developed a loop-mediated isothermal amplification (LAMP)-based electrochemical test for the detection of SARS-CoV-2 that causes COVID-19. The test is based on the oxidation-reduction reaction between pyrophosphates (generated from positive LAMP reaction) and molybdate that is detected by cyclic voltammetry using inexpensive and disposable carbon screen printed electrodes. Our test showed higher sensitivity (detecting as low as 5.29 RNA copies/μL) compared to the conventional fluorescent reverse transcriptase (RT)-LAMP. We validated our tests using human serum and saliva spiked with SARS-CoV-2 RNA and clinical (saliva and nasal-pharyngeal) swab samples demonstrating 100% specificity and 93.33% sensitivity. Our assay provides a rapid, specific, and sensitive test with an electrochemical readout in less than 45 min that could be adapted for point-of-care settings.
为了控制像新冠疫情这样给人类健康和全球经济带来巨大负担的疾病爆发，需要快速、可靠、灵敏、便携且准确的诊断方法。在此，我们开发了一种基于环介导等温扩增（LAMP）的电化学检测方法，用于检测引发新冠疫情的严重急性呼吸综合征冠状病毒2（SARS-CoV-2）。该检测方法基于焦磷酸（由阳性LAMP反应产生）与钼酸盐之间的氧化还原反应，通过使用廉价的一次性碳丝网印刷电极的循环伏安法进行检测。与传统的荧光逆转录（RT）-LAMP相比，我们的检测方法具有更高的灵敏度（低至可检测5.29个RNA拷贝/微升）。我们使用添加了SARS-CoV-2 RNA的人血清和唾液以及临床（唾液和鼻咽）拭子样本对检测方法进行了验证，结果显示其特异性为100%，灵敏度为93.33%。我们的检测方法可在不到45分钟内提供具有电化学读数的快速、特异且灵敏的检测，适用于即时检测场景。
