Objectives: Development and validation of a single-step and accurate reverse transcriptase loop-mediated isothermal amplification technique (RT-LAMP) for rapid identification of SARS-CoV-2 relative to commercial quantitative reverse transcriptase real-time PCR (qRT-PCR) assays to allow prompt initiation of proper medical care and containment of virus spread.

Methods: Primers showing optimal in-silico features were subjected to analytical sensitivity and specificity to assess the limit of detection (LOD) and cross-reaction with closely- and distantly-related viral species, and clinically prominent bacterial and fungal species. In order to evaluate the clinical utility, our RT-LAMP was subjected to a large number of clinical samples, including 213 negative and 47 positive patients, relative to two commercial quantitative RT-PCR assays.

Results: The analytical specificity and sensitivity of our assay was 100% and 500 copies/ml when serial dilution was performed in both water and sputum. Subjecting our RT-LAMP assay to clinical samples showed a high degree of specificity (99.5%), sensitivity (91.4%), positive predictive value (97.7%), and negative predictive value (98.1%) when used relative to qRT-PCR. Our RT-LAMP assay was two times faster than qRT-PCR and is storable at room temperature. A suspected case that later became positive tested positive using both our RT-LAMP and the two qRT-PCR assays, which shows the capability of our assay for screening purposes.

Conclusions: We present a rapid RT-LAMP assay that could extend the capacity of laboratories to process two times more clinical samples relative to qRT-PCR and potentially could be used for high-throughput screening purposes when demand is increasing at critical situations.

目标： 一步和准确的逆转录酶环介导的等温扩增技术（RT-LAMP）的开发和验证，可相对于商业定量逆转录酶实时PCR（qRT-PCR）分析快速鉴定SARS-CoV-2立即开始适当的医疗护理并控制病毒传播。

方法： 引物显示最佳 电脑内对这些特征进行分析敏感性和特异性，以评估检测限（LOD）和与近缘和远缘相关病毒物种以及临床上著名的细菌和真菌物种的交叉反应。为了评估临床效用，相对于两种商业定量RT-PCR分析，我们的RT-LAMP接受了大量临床样品，包括213例阴性和47例阳性患者。

结果：当在水和痰液中进行系列稀释时，我们测定的分析特异性和灵敏度为100％和500拷贝/ ml。与qRT-PCR相比，将我们的RT-LAMP分析应用于临床样品显示出高度的特异性（99.5％），敏感性（91.4％），阳性预测值（97.7％）和阴性预测值（98.1％）。我们的RT-LAMP分析比qRT-PCR快两倍，并且可以在室温下保存。使用我们的RT-LAMP和两个qRT-PCR分析法，后来都变成阳性的可疑病例被测试为阳性，这表明我们的分析方法具有筛查目的。

结论： 我们提出了一种快速的RT-LAMP分析方法，它可以将实验室的处理能力扩展到相对于qRT-PCR两倍的临床样品，并且可以在关键情况下需求增加时用于高通量筛选。