A one-step real-time RT-PCR assay for simultaneous typing of SARS-CoV-2 mutations associated with the E484K and N501Y spike protein amino-acid substitutions.
A one-step real-time RT-PCR assay for simultaneous typing of SARS-CoV-2 mutations associated with the E484K and N501Y spike protein amino-acid substitutions.
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DOI：
10.1016/j.jviromet.2021.114242
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发表时间：
2021-10
影响因子：
3.1
通讯作者：
Dovas CI
中科院分区：
文献类型：
--
作者：
Chaintoutis SC;Chassalevris T;Tsiolas G;Balaska S;Vlatakis I;Mouchtaropoulou E;Siarkou VI;Tychala A;Koutsioulis D;Skoura L;Argiriou A;Dovas CI
SARS-CoV-2突变的出现导致S蛋白氨基酸替换N501Y和E484K，这两种氨基酸分别与增强的传播性和免疫逃逸有关，需要立即采取行动，因此快速识别它们至关重要。为了同时分型这两种关注突变（MOCs），我们开发了一种采用4个锁定核酸（LNA）修饰的TaqMan探针的一步实时RT-PCR检测方法。该方法高度敏感，LOD为117拷贝/反应，扩增效率为bb0 94%，线性范围超过5 log10拷贝/反应。利用人类和动物中已知的sars - cov -2阳性和阴性样本对该方法进行验证，结果表明该方法能够正确识别野生型菌株以及具有一个或两个靶向氨基酸取代的菌株，从而成为快速鉴定MOC的有用预筛选工具。为了便于快速设计能够检测新出现的moc的类似检测方法，解释了开发该检测方法的基本原理。
The emergence of SARS-CoV-2 mutations resulting in the S protein amino-acid substitutions N501Y and E484K, which have been associated with enhanced transmissibility and immune escape, respectively, necessitates immediate actions, for which their rapid identification is crucial. For the simultaneous typing of both of these mutations of concern (MOCs), a one-step real-time RT-PCR assay employing four locked nucleic acid (LNA) modified TaqMan probes was developed. The assay is highly sensitive with a LOD of 117 copies/reaction, amplification efficiencies >94 % and a linear range of over 5 log10 copies/reaction. Validation of the assay using known SARS-CoV-2-positive and negative samples from human and animals revealed its ability to correctly identify wild type strains, and strains possessing either one or both targeted amino-acid substitutions, thus comprising a useful pre-screening tool for rapid MOC identification. The basic principles of the methodology for the development of the assay are explained in order to facilitate the rapid design of similar assays able to detect emerging MOCs.
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影响因子：
2.9
作者：
Owczarzy, Richard;You, Yong;Groth, Christopher L.;Tataurov, Andrey V.
通讯作者：
Tataurov, Andrey V.
DOI：
10.1016/j.dental.2021.01.015
发表时间：
2021-03
期刊：
Dental materials : official publication of the Academy of Dental Materials
影响因子：
--
作者：
Durner J;Burggraf S;Czibere L;Tehrani A;Watts DC;Becker M
通讯作者：
Becker M
影响因子：
13.2
作者：
Chan, Jasper Fuk-Woo;Kok, Kin-Hang;Yuen, Kwok-Yung
通讯作者：
Yuen, Kwok-Yung
LNA探针的设计可改善不匹配歧视。
影响因子：
14.9
作者：
You, Yong;Moreira, Bernardo G.;Behlke, Mark A.;Owczarzy, Richard
通讯作者：
Owczarzy, Richard
影响因子：
4.3
作者：
Chaintoutis, Serafeim C.;Siarkou, Victoria I.;Dovas, Chrysostomos I.
通讯作者：
Dovas, Chrysostomos I.