RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2
RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2
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DOI：
10.1111/1751-7915.13586
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发表时间：
2020-04-25
影响因子：
5.7
通讯作者：
Cui, Zhanfeng
中科院分区：
文献类型：
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作者：
Huang, Wei E.;Lim, Boon;Cui, Zhanfeng
全球大流行的冠状病毒SARS-CoV-2已导致大量感染人群患上COVID-19。为了遏制病毒的传播，世卫组织迫切要求扩大筛查和检测范围;因此，需要一种快速简便的诊断方法。我们设计了4对引物，每组6对，分别针对SARS CoV-2病毒RNA的orf 1ab、S基因和N基因区域，采用逆转录环介导等温扩增（RT-LAMP）技术，在30 min内实现对SARS CoV-2的检测。使用比色变化来报告结果，这使得病毒RNA扩增的结果能够通过肉眼读取，而不需要昂贵或专用的仪器。灵敏度可以是样品中每ml 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个阳性和8个阴性。检测结果与常规RT-qPCR一致。此外，我们还表明，无需RNA提取的一步法直接从样品中实现RNA扩增是可行的。这种快速、简单和灵敏的RT-LAMP方法为在公共领域和医院，特别是农村地区的区域医院和医疗中心进行大规模筛查铺平了道路。
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.