Essential properties and pitfalls of colorimetric Reverse Transcription Loop-mediated Isothermal Amplification as a point-of-care test for SARS-CoV-2 diagnosis.
Essential properties and pitfalls of colorimetric Reverse Transcription Loop-mediated Isothermal Amplification as a point-of-care test for SARS-CoV-2 diagnosis.
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DOI：
10.1186/s10020-021-00289-0
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发表时间：
2021-03-26
期刊：
影响因子：
--
通讯作者：
Blanes L
中科院分区：
文献类型：
--
作者：
de Oliveira Coelho B;Sanchuki HBS;Zanette DL;Nardin JM;Morales HMP;Fornazari B;Aoki MN;Blanes L
SARS-CoV-2逆转录环介导等温扩增(RT-LAMP)比色法检测SARS-CoV-2是一种灵敏而特异的护理点分子生物学技术，仅需30分钟即可检测到病毒。在这篇手稿中，我们描述了一些文献中仍未正确描述的技术的细微差别：三种颜色簇的存在；病毒载量与颜色变化的相关性；以及使用内部对照避免假阴性结果的重要性。为了实现这些发现，我们对466例SARS-CoV-2 RT-qPCR验证的临床样本进行了比色RT-LAMP分析，颜色定量分别在434 nm和560 nm处测量。首先，我们确定敏感性为93.8%，特异性为90.4%。除了粉色(阴性)和黄色(阳性)产生的颜色外，我们首次报告了可能导致错误诊断的橙色簇的存在。我们还使用RT-qPCR和RT-LAMP证明了低病毒载量与 >； 30的CT值相关，导致橙色。我们还证明，直到出现症状的第五天，病毒载量仍然相对较高的时候，比色RT-LAMP对新冠肺炎的诊断都是有效的。本研究报告了用于SARS-CoV-2诊断的比色法RT-LAMP的特性和适应症，减少了错误结果、解释，并优化了分子诊断测试的应用。在线版本包含补充材料，可在10.1186/s10020-021-00289-0上查阅。
SARS-CoV-2 Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) colorimetric detection is a sensitive and specific point-of-care molecular biology technique used to detect the virus in only 30 min. In this manuscript we have described a few nuances of the technique still not properly described in the literature: the presence of three colors clusters; the correlation of the viral load with the color change; and the importance of using an internal control to avoid false-negative results. To achieve these findings, we performed colorimetric RT-LAMP assays of 466 SARS-CoV-2 RT-qPCR validated clinical samples, with color quantification measured at 434 nm and 560 nm. First we determinate a sensitivity of 93.8% and specificity of 90.4%. In addition to the pink (negative) and yellow (positive) produced colors, we report for the first time the presence of an orange color cluster that may lead to wrong diagnosis. We also demonstrated using RT-qPCR and RT-LAMP that low viral loads are related to Ct values > 30, resulting in orange colors. We also demonstrated that the diagnosis of COVID-19 by colorimetric RT-LAMP is efficient until the fifth symptoms day when the viral load is still relatively high. This study reports properties and indications for colorimetric RT-LAMP as point-of-care for SARS-CoV-2 diagnostic, reducing false results, interpretations and optimizing molecular diagnostics tests application. The online version contains supplementary material available at 10.1186/s10020-021-00289-0.
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影响因子：
3.1
作者：
Howson ELA;Kidd SP;Armson B;Goring A;Sawyer J;Cassar C;Cross D;Lewis T;Hockey J;Rivers S;Cawthraw S;Banyard A;Anderson P;Rahou S;Andreou M;Morant N;Clark D;Walsh C;Laxman S;Houghton R;Slater-Jefferies J;Costello P;Brown I;Cortes N;Godfrey KM;Fowler VL
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作者：
Haguet, Helene;Douxfils, Jonathan;Favresse, Julien
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Favresse, Julien
DOI：
10.1007/s10096-020-03913-9
发表时间：
2020-04-27
影响因子：
4.5
作者：
La Scola, Bernard;Le Bideau, Marion;Raoult, Didier
通讯作者：
Raoult, Didier
DOI：
10.1073/pnas.2011221117
发表时间：
2020-09-29
影响因子：
11.1
作者：
Rabe BA;Cepko C
通讯作者：
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