Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction.
Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction.
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DOI：
10.1038/s41598-021-81487-y
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发表时间：
2021-01-28
影响因子：
4.6
通讯作者：
Williams Z
中科院分区：
文献类型：
--
作者：
Wei S;Kohl E;Djandji A;Morgan S;Whittier S;Mansukhani M;Hod E;D'Alton M;Suh Y;Williams Z
COVID-19 大流行导致迫切需要能够在全球范围内迅速扩展的快速护理诊断测试。我们开发并测试了一种基于逆转录环介导等温扩增 (RT-LAMP) 的高度灵敏且稳健的检测方法，该检测方法使用现成的试剂和简单的加热块，并使用人为的掺入和实际临床样本。对 RNA 加标样品进行的 RT-LAMP 测试显示，病毒运输介质的检测限 (LoD) 为 2.5 拷贝/μl。直接对病毒转运介质中的临床鼻咽拭子样本进行 RT-LAMP 检测，阳性百分比一致性 (PPA) (17/20) 为 85%，阴性百分比一致性 (NPV) 为 100%，并在 30 分钟内得出结果。我们基于 RT-LAMP 的优化测试方法是一个可扩展的系统，该系统足够灵敏且稳健，可以在护理点 30 分钟内直接对病毒运输介质中的临床鼻咽拭子样本进行 SARS-CoV-2 测试，而无需专门或专有的设备或试剂。这种经济高效的一步式检测方法可随时用于全球范围内的 COVID-19 检测，特别是在资源匮乏的环境中。
The COVID-19 pandemic has resulted in an urgent need for a rapid, point of care diagnostic testing that could be rapidly scaled on a worldwide level. We developed and tested a highly sensitive and robust assay based on reverse transcription loop mediated isothermal amplification (RT-LAMP) that uses readily available reagents and a simple heat block using contrived spike-in and actual clinical samples. RT-LAMP testing on RNA-spiked samples showed a limit of detection (LoD) of 2.5 copies/μl of viral transport media. RT-LAMP testing directly on clinical nasopharyngeal swab samples in viral transport media had an 85% positive percentage agreement (PPA) (17/20), and 100% negative percentage agreement (NPV) and delivered results in 30 min. Our optimized RT-LAMP based testing method is a scalable system that is sufficiently sensitive and robust to test for SARS-CoV-2 directly on clinical nasopharyngeal swab samples in viral transport media in 30 min at the point of care without the need for specialized or proprietary equipment or reagents. This cost-effective and efficient one-step testing method can be readily available for COVID-19 testing world-wide, especially in resource poor settings.
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