Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China.
Department of Engineering Science, University of Oxford, Parks Road, OX1 3PJ, Oxford, UK.
Microb Biotechnol. 2020 Jul;13(4):950-961. doi: 10.1111/1751-7915.13586. Epub 2020 Apr 25.
The pandemic coronavirus SARS-CoV-2 in the world has caused a large infected population suffering from COVID-19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) to achieve the detection of SARS-CoV-2 in 30 min. We designed four sets of LAMP primers (6 primers in each set), targeting the viral RNA of SARS-CoV-2 in the regions of orf1ab, S gene and N gene. A colorimetric change was used to report the results, which enables the outcome of viral RNA amplification to be read by the naked eye without the need of expensive or dedicated instrument. The sensitivity can be 80 copies of viral RNA per ml in a sample. We validated the RT-LAMP method in a hospital in China, employing 16 clinic samples with 8 positives and 8 negatives. The testing results are consistent with the conventional RT-qPCR. In addition, we also show that one-step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. This rapid, simple and sensitive RT-LAMP method paves a way for a large screening at public domain and hospitals, particularly regional hospitals and medical centres in rural areas.
在世界范围内流行的冠状病毒 SARS-CoV-2 导致了大量感染 COVID-19 的人群。为了遏制病毒的传播,世界卫生组织紧急要求扩大筛查和检测范围;因此,需要一种快速简单的诊断方法。我们应用逆转录环介导等温扩增(RT-LAMP)在 30 分钟内实现了对 SARS-CoV-2 的检测。我们设计了四组 LAMP 引物(每组 6 个引物),针对 SARS-CoV-2 的病毒 RNA 在orf1ab、S 基因和 N 基因区域。通过颜色变化来报告结果,这使得无需昂贵或专用仪器即可通过肉眼读取病毒 RNA 扩增的结果。该方法的灵敏度可达到样本中每毫升 80 个病毒 RNA 拷贝。我们在中国的一家医院验证了 RT-LAMP 方法,使用了 16 个临床样本,其中 8 个阳性,8 个阴性。检测结果与常规 RT-qPCR 一致。此外,我们还表明,一步法无需提取 RNA 即可直接从样本中扩增 RNA。这种快速、简单、灵敏的 RT-LAMP 方法为在公共场所和医院,特别是在农村地区的地区医院和医疗中心进行大规模筛查铺平了道路。