A novel One-pot rapid diagnostic technology for COVID-19.
A novel One-pot rapid diagnostic technology for COVID-19.
复制标题
一种新的新冠肺炎一锅快速诊断技术。
DOI：
10.1016/j.aca.2021.338310
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发表时间：
2021-04-15
影响因子：
6.2
通讯作者：
Li S
中科院分区：
文献类型：
--
作者：
Li J;Hu X;Wang X;Yang J;Zhang L;Deng Q;Zhang X;Wang Z;Hou T;Li S
由SARS-CoV-2引起的新型冠状病毒病（COVID-19）是一种持续的全球大流行病，发病率和死亡率很高。RT-qPCR已成为大多数国家检测SARS-CoV-2的诊断标准。COVID-19的诊断通常依赖于RT-qPCR介导的SARS-CoV-2病毒RNA的鉴定，这是昂贵的，劳动密集的，并且需要专门的培训和设备。在此，我们建立了一种基于甲酰胺和比色RT-LAMP的新型单管快速诊断方法（One-Pot RT-LAMP），可用于诊断COVID-19，而无需提取特异性病毒RNA。该技术可在45 min内目视检测SARS-CoV-2，在提取的RNA中检测限为5拷贝/反应，在病毒转运培养基中检测限约为7.66拷贝/μL。结果表明，该方法特异性高，与SARS冠状病毒、MERS冠状病毒、人感染性流感病毒（甲型H1 N1/H3 N2、B型）等12种病毒无交叉反应。我们通过成功用于45份临床鼻咽拭子样本的分析来验证这种一锅法RT-LAMP方法，产生与传统RT-qPCR分析相同的结果，同时相对于商业RT-qPCR方法实现了良好的选择性和灵敏度。因此，该One-Pot RT-LAMP技术可能是进行高灵敏度、低成本和快速鉴定SARS-CoV-2的有效手段，而无需提取病毒RNA。
Novel coronavirus disease (COVID-19) caused by SARS-CoV-2 is an ongoing global pandemic associated with high rates of morbidity and mortality. RT-qPCR has become the diagnostic standard for the testing of SARS-CoV-2 in most countries. COVID-19 diagnosis generally relies upon RT-qPCR-mediated identification of SARS-CoV-2 viral RNA, which is costly, labor-extensive, and requires specialized training and equipment. Herein, we established a novel one-tube rapid diagnostic approach based upon formamide and colorimetric RT-LAMP (One-Pot RT-LAMP) that can be used to diagnose COVID-19 without the extraction of specific viral RNA. The technique could visually detect SARS-CoV-2 within 45 min with a limit of detection of 5 copies per reaction in extracted RNA, and about 7.66 virus copies per μL in viral transport medium. The One-Pot RT-LAMP test showed a high specificity without cross-reactivity with 12 viruses including SARS-CoV, MERS-CoV, and human infectious influenza virus (H1N1/H3N2 of influenza A and B virus, ect. We validated this One-Pot RT-LAMP approach by its successful use for the analysis of 45 clinical nasopharyngeal swab samples, yielding results identical to those of traditional RT-qPCR analyses, while achieving good selectivity and sensitivity relative to a commercial RT-qPCR approach. As such, this One-Pot RT-LAMP technology may be a valid means of conducting high-sensitivity, low-cost and rapid SARS-CoV-2 identification without the extraction of viral RNA.
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DOI：
10.1073/pnas.2014739117
发表时间：
2020-09-15
影响因子：
11.1
作者：
Ganguli A;Mostafa A;Berger J;Aydin MY;Sun F;Ramirez SAS;Valera E;Cunningham BT;King WP;Bashir R
通讯作者：
Bashir R
影响因子：
16.6
作者：
Ding X;Yin K;Li Z;Lalla RV;Ballesteros E;Sfeir MM;Liu C
通讯作者：
Liu C
影响因子：
28.3
作者：
Gorbalenya, Alexander E.;Baker, Susan C.;Ziebuhr, John
通讯作者：
Ziebuhr, John
影响因子：
5.7
作者：
Huang, Wei E.;Lim, Boon;Cui, Zhanfeng
通讯作者：
Cui, Zhanfeng
影响因子：
9.3
作者：
Behrmann, Ole;Bachmann, Iris;Hufert, Frank T.
通讯作者：
Hufert, Frank T.