Fast, reliable and easy to handle methods are required to facilitate urgently needed point-of-care testing (POCT) in the current coronavirus pandemic. Life-threatening severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly spread all over the world, infecting more than 33,500,000 people and killing over 1 million of them as of October 2020. Infected individuals without any symptoms might still transfer the virus to others underlining the extraordinary transmissibility of this new coronavirus. In order to identify early infections effectively, treat patients on time and control disease spreading, rapid, accurate and onsite testing methods are urgently required. Here we report the development of a loop-mediated isothermal amplification (LAMP) based method to detect SARS-CoV-2 genes ORF8 and N directly from pharyngeal swab samples. The established reverse transcription LAMP (RT-LAMP) assay detects SARS-CoV-2 directly from pharyngeal swab samples without previous time-consuming and laborious RNA extraction. The assay is sensitive and highly specific for SARS-CoV-2 detection, showing no cross reactivity when tested on 20 other respiratory pathogens. The assay is 12 times faster and 10 times cheaper than routine reverse transcription real-time polymerase chain reaction, depending on the assay used. The fast and easy to handle RT-LAMP assay amplifying specifically the genomic regions ORF8 and N of SARS-CoV-2 is ideally suited for POCT at e.g. railway stations, airports or hospitals. Given the current pandemic situation, rapid, cost efficient and onsite methods like the here presented RT-LAMP assay are urgently needed to contain the viral spread.

中文翻译：

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使用逆转录环介导的等温扩增（RT-LAMP）快速即时检测SARS-CoV-2

需要快速，可靠且易于处理的方法来促进当前冠状病毒大流行中迫切需要的即时检验（POCT）。到2020年10月，威胁生命的严重急性呼吸综合征冠状病毒2（SARS-CoV-2）在世界范围内迅速传播，感染了33,500,000多人，并杀死了超过100万人。病毒对其他人的感染，突显了这种新型冠状病毒的非凡传播性。为了有效地识别早期感染，及时治疗患者并控制疾病传播，迫切需要快速，准确和现场的检测方法。在这里，我们报告基于环介导的等温扩增（LAMP）的方法的发展，以直接从咽拭子样本中检测SARS-CoV-2基因ORF8和N。已建立的逆转录LAMP（RT-LAMP）分析法可直接从咽拭子样本中检测SARS-CoV-2，而无需耗时费力的RNA提取。该测定法对SARS-CoV-2检测灵敏且具有高度特异性，对其他20种呼吸道病原体进行测试时，无交叉反应。该测定比常规逆转录实时聚合酶链反应快12倍，便宜10倍，具体取决于所使用的测定。快速且易于操作的RT-LAMP测定法特别扩增了SARS-CoV-2的基因组区域ORF8和N，非常适合于例如火车站，机场或医院的POCT。考虑到当前的大流行情况，迫切需要一种快速，经济高效的现场方法，如此处介绍的RT-LAMP分析，以遏制病毒传播。