Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin.
Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin.
复制标题
使用螯合树脂进行灵敏的免提取SARS-CoV-2 RNA病毒检测。
DOI：
10.1016/j.isci.2021.102960
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发表时间：
2021-09-24
期刊：
影响因子：
5.8
通讯作者：
Hufnagel RB
中科院分区：
文献类型：
--
作者：
Guan B;Frank KM;Maldonado JO;Beach M;Pelayo E;Warner BM;Hufnagel RB
目前SARS-CoV-2的常规检测包括用鼻咽拭子收集患者样本，在病毒转运培养基中转运期间储存拭子，提取RNA和定量逆转录PCR（RT-qPCR）。我们开发了一种使用螯合树脂Chelex的简化制备方法，该方法避免了病毒检测期间的RNA提取。将直接检测RT-qPCR和数字液滴PCR与目前常规的RNA提取方法进行了模拟样本和患者标本的比较。与单独加热相比，在Chelex存在下的热处理显著提高了检测灵敏度，并且缺乏RNA提取缩短了整体诊断工作流程。此外，初始样品加热步骤灭活SARS-CoV-2感染性，从而提高了工作流程的安全性。这种快速RNA制备和检测方法适用于各种样品，对检测人员安全，适用于高通量平台上的标准临床采集和检测。Chelex树脂在普通缓冲液中保存SARS-CoV-2 RNA，允许直接RT-qPCR Chelex方法为RNA病原体提供了一种安全、经济和敏感的检测方法生物科学;分子生物学实验方法;病毒学;生物科学研究方法;
Current conventional detection of SARS-CoV-2 involves collection of a patient’s sample with a nasopharyngeal swab, storage of the swab during transport in a viral transport medium, extraction of RNA, and quantitative reverse transcription PCR (RT-qPCR). We developed a simplified preparation method using a chelating resin, Chelex, which obviates RNA extraction during viral testing. Direct detection RT-qPCR and digital droplet PCR were compared to the current conventional method with RNA extraction for simulated samples and patient specimens. The heat treatment in the presence of Chelex markedly improved detection sensitivity as compared to heat alone, and lack of RNA extraction shortens the overall diagnostic workflow. Furthermore, the initial sample heating step inactivates SARS-CoV-2 infectivity, thus improving workflow safety. This fast RNA preparation and detection method is versatile for a variety of samples, safe for testing personnel, and suitable for standard clinical collection and testing on high-throughput platforms. The COVID-19 pandemic caused supply shortages for diagnostic tests Chelex resin preserves SARS-CoV-2 RNA in common buffers allowing direct RT-qPCR The Chelex method presents a safe, economic, and sensitive test for RNA pathogens Biological sciences; Molecular biology experimental approach; Virology; Biological sciences research methodologies;
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影响因子：
16.6
作者：
Smyrlaki I;Ekman M;Lentini A;Rufino de Sousa N;Papanicolaou N;Vondracek M;Aarum J;Safari H;Muradrasoli S;Rothfuchs AG;Albert J;Högberg B;Reinius B
通讯作者：
Reinius B
影响因子：
2.7
作者：
WALSH, PS;METZGER, DA;HIGUCHI, R
通讯作者：
HIGUCHI, R
影响因子：
9.4
作者：
Barat B;Das S;De Giorgi V;Henderson DK;Kopka S;Lau AF;Miller T;Moriarty T;Palmore TN;Sawney S;Spalding C;Tanjutco P;Wortmann G;Zelazny AM;Frank KM
通讯作者：
Frank KM
DOI：
10.1016/j.jcv.2020.104423
发表时间：
2020-07-01
期刊：
Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
影响因子：
--
作者：
Merindol, Natacha;Pepin, Genevieve;Danylo, Alexis
通讯作者：
Danylo, Alexis
影响因子：
8.8
作者：
Chu, Allen Wing-Ho;Chan, Wan-Mui;Kai-Wang To, Kelvin
通讯作者：
Kai-Wang To, Kelvin