The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak urgently necessitates sensitive and convenient COVID-19 diagnostics for the containment and timely treatment of patients. We aimed to develop and validate a novel reverse transcription–loop-mediated isothermal amplification (RT-LAMP) assay to detect SARS-CoV-2. Patients with suspected COVID-19 and close contacts were recruited from two hospitals between 26 January and 8 April 2020. Respiratory samples were collected and tested using RT-LAMP, and the results were compared with those obtained by reverse transcription-quantitative PCR (RT-qPCR). Samples yielding inconsistent results between these two methods were subjected to next-generation sequencing for confirmation. RT-LAMP was also applied to an asymptomatic COVID-19 carrier and patients with other respiratory viral infections. Samples were collected from a cohort of 129 cases (329 nasopharyngeal swabs) and an independent cohort of 76 patients (152 nasopharyngeal swabs and sputum samples). The RT-LAMP assay was validated to be accurate (overall sensitivity and specificity of 88.89% and 99.00%, respectively) and diagnostically useful (positive and negative likelihood ratios of 88.89 and 0.11, respectively). RT-LAMP showed increased sensitivity (88.89% versus 81.48%) and high consistency (kappa, 0.92) compared to those of RT-qPCR for SARS-CoV-2 screening while requiring only constant-temperature heating and visual inspection. The time required for RT-LAMP was less than 1 h from sample preparation to the result. In addition, RT-LAMP was feasible for use with asymptomatic patients and did not cross-react with other respiratory pathogens. The developed RT-LAMP assay offers rapid, sensitive, and straightforward detection of SARS-CoV-2 infection and may aid the expansion of COVID-19 testing in the public domain and hospitals.

中文翻译：

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用于 SARS-CoV-2 感染的快速灵敏的环路介导等温扩增试验的开发和临床应用。

严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 的爆发迫切需要灵敏、方便的 COVID-19 诊断方法，以遏制和及时治疗患者。我们旨在开发和验证一种新型逆转录环介导等温扩增 (RT-LAMP) 检测方法来检测 SARS-CoV-2。2020 年 1 月 26 日至 4 月 8 日期间，从两家医院招募疑似 COVID-19 的患者和密切接触者。收集呼吸道样本并使用 RT-LAMP 进行检测，并将结果与​​逆转录定量 PCR（RT- qPCR）。对这两种方法产生不一致结果的样品进行下一代测序以进行确认。RT-LAMP 还应用于无症状的 COVID-19 携带者和其他呼吸道病毒感染的患者。从 129 例病例（329 份鼻咽拭子）和 76 名患者（152 份鼻咽拭子和痰液样本）的独立队列中收集样本。RT-LAMP 检测被证实是准确的（总体敏感性和特异性分别为 88.89% 和 99.00%）和诊断有用（阳性和阴性似然比分别为 88.89 和 0.11）。与用于 SARS-CoV-2 筛查的 RT-qPCR 相比，RT-LAMP 显示出更高的灵敏度（88.89% 对 81.48%）和高一致性（kappa，0.92），同时只需要恒温加热和目视检查。RT-LAMP 从样品制备到获得结果所需的时间不到 1 小时。此外，RT-LAMP 可用于无症状患者，并且不会与其他呼吸道病原体发生交叉反应。开发的 RT-LAMP 检测提供了对 SARS-CoV-2 感染的快速、灵敏和直接的检测，并可能有助于在公共领域和医院中扩展 COVID-19 检测。