The ongoing outbreaks of the coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have resulted in unprecedented challenges to global health. To effectively contain the COVID-19 transmission, rapid tests for detecting existing SARS-CoV-2 infections and assessing virus spread are critical. To address the huge need for ever-increasing tests, we developed a facile all-in-one nucleic acid testing assay by combining Si-OH activated glass bead (aGB)-based viral RNA fast extraction and in situ colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) detection in a single tube. aGBs demonstrate a strong ability to capture viral RNA in a guanidinium-based lysis buffer, and the purified aGBs/RNA composite, without RNA elution step, could be directly used to perform RT-LAMP assay. The assay was well characterized by using a novel SARS-CoV-2-like coronavirus GX/P2V, and showed a limit of detection (LOD) of 15 copies per μL in simulated clinical samples within 50 min. We further demonstrated our assay by testing simulated SARS-CoV-2 pseudovirus samples, showing an LOD of 32 copies per μL and high specificity without cross-reactivity with the most closely related GX/P2V or host DNA/RNA. The all-in-one approach developed in this study has the potential as a simple, scalable, and time-saving alternative for point-of-care testing of SARS-CoV-2 in low-income regions, as well as a promising tool for at-home testing.

中文翻译：

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用于 SARS-CoV-2 即时检测的多合一原位比色 RT-LAMP 检测

由严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 引起的 2019 年冠状病毒病 (COVID-19) 大流行的持续爆发给全球健康带来了前所未有的挑战。为了有效遏制 COVID-19 传播，检测现有 SARS-CoV-2 感染和评估病毒传播的快速测试至关重要。为了满足不断增加的测试的巨大需求，我们通过结合基于 Si-OH 活化玻璃珠 (aGB) 的病毒 RNA 快速提取和原位检测，开发了一种简便的一体化核酸检测方法在单管中进行比色逆转录环介导的等温扩增 (RT-LAMP) 检测。aGBs 表现出在基于胍的裂解缓冲液中捕获病毒 RNA 的强大能力，并且纯化的 aGBs/RNA 复合物无需 RNA 洗脱步骤，可直接用于进行 RT-LAMP 检测。该测定通过使用新型 SARS-CoV-2 样冠状病毒 GX/P2V 进行了很好的表征，并在 50 分钟内在模拟临床样本中显示检测限 (LOD) 为每微升 15 个拷贝。我们通过测试模拟的 SARS-CoV-2 假病毒样本进一步证明了我们的分析，显示出每微升 32 个拷贝的 LOD 和高特异性，与最密切相关的 GX/P2V 或宿主 DNA/RNA 没有交叉反应。本研究中开发的多合一方法具有作为一种简单、可扩展、