A Simple, Affordable, Rapid, Stabilized, Colorimetric, Versatile RT-LAMP Assay to Detect SARS-CoV-2.
A Simple, Affordable, Rapid, Stabilized, Colorimetric, Versatile RT-LAMP Assay to Detect SARS-CoV-2.
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DOI：
10.3390/diagnostics11030438
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发表时间：
2021-03-04
期刊：
影响因子：
--
通讯作者：
Muro A
中科院分区：
文献类型：
--
作者：
García-Bernalt Diego J;Fernández-Soto P;Domínguez-Gil M;Belhassen-García M;Bellido JLM;Muro A
SARS-CoV-2大流行迫使全球所有国家迅速开发和实施广泛的检测，以控制和管理冠状病毒病2019年(新冠肺炎)。逆转录(RT)-qPCR是新冠肺炎的金标准分子诊断方法，主要用于自动化测试平台。这些系统准确有效，但也昂贵、耗时、高科技、依赖基础设施，目前正面临商业试剂供应短缺的问题。逆转录-环介导的等温扩增(RT-LAMP)可以作为一种替代检测方法。在这里，我们提出了一种新型的通用(实时和比色)RT-LAMP，用于针对新型病毒基因组的ORF1ab和N基因的简单(一步)、负担得起(~1.7欧元/样本)和快速检测SARS-CoV-2。我们对RT-qPCR阳性的临床样本进行了检测，在25分钟内获得了最大的阳性结果。此外，还开发了一种新的30分钟一步干燥方案来稳定RT-LAMP反应混合物，允许它们在室温下功能性储存长达两个月，正如Q10所预测的那样。这种干式RT-LAMP方法适用于潜在的即用型新冠肺炎诊断。在进一步测试和验证之后，它可以很容易地应用于发达国家和低收入国家，产生快速和可靠的结果。
The SARS-CoV-2 pandemic has forced all countries worldwide to rapidly develop and implement widespread testing to control and manage the Coronavirus Disease 2019 (COVID-19). reverse-transcription (RT)-qPCR is the gold standard molecular diagnostic method for COVID-19, mostly in automated testing platforms. These systems are accurate and effective, but also costly, time-consuming, high-technological, infrastructure-dependent, and currently suffer from commercial reagent supply shortages. The reverse-transcription loop-mediated isothermal amplification (RT-LAMP) can be used as an alternative testing method. Here, we present a novel versatile (real-time and colorimetric) RT-LAMP for the simple (one-step), affordable (~1.7 €/sample), and rapid detection of SARS-CoV-2 targeting both ORF1ab and N genes of the novel virus genome. We demonstrate the assay on RT-qPCR-positive clinical samples, obtaining most positive results under 25 min. In addition, a novel 30-min one-step drying protocol has been developed to stabilize the RT-LAMP reaction mixtures, allowing them to be stored at room temperature functionally for up to two months, as predicted by the Q10. This Dry-RT-LAMP methodology is suitable for potentially ready-to-use COVID-19 diagnosis. After further testing and validation, it could be easily applied both in developed and in low-income countries yielding rapid and reliable results.
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影响因子：
4.6
作者：
Garcia-Bernalt Diego, J.;Fernandez-Soto, P.;Muro, A.
通讯作者：
Muro, A.
影响因子：
6.1
作者：
Liu, Kui;Fang, Yuan-Yuan;Liu, Hui-Guo
通讯作者：
Liu, Hui-Guo
开发和验证快速、单步逆转录酶环介导的等温扩增 (RT-LAMP) 系统，有望用于可靠和高通量的 COVID-19 筛查
影响因子：
5.7
作者：
Jiang, Ming-hua;Pan, Wei-hua;Li, X. J.
通讯作者：
Li, X. J.
影响因子：
168.9
作者：
Huang, Chaolin;Wang, Yeming;Cao, Bin
通讯作者：
Cao, Bin
影响因子：
5.7
作者：
Huang, Wei E.;Lim, Boon;Cui, Zhanfeng
通讯作者：
Cui, Zhanfeng