Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR.
Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR.
复制标题
DOI：
10.1038/s41467-020-18611-5
复制
发表时间：
2020-09-23
影响因子：
16.6
通讯作者：
Reinius B
中科院分区：
文献类型：
--
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
由严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）引起的2019冠状病毒病（COVID-19）通常通过逆转录聚合酶链反应（RT-PCR）检测患者样本中的病毒RNA进行诊断，但RNA提取构成了当前检测的主要瓶颈。方法的简化可以提高诊断的可用性和效率，有利于病人护理和感染控制。在这里，我们描述了在COVID-19检测中通过直接对热灭活或裂解样本进行RT-PCR来避免RNA提取的方法。我们的数据，包括使用597个临床患者样本和标准化诊断系统的基准测试，表明直接RT-PCR是基于提取的测试的可行选择。使用受控量的活性SARS-CoV-2，我们通过空斑试验确认热灭活的有效性，并评估各种通用缓冲液作为直接RT-PCR的运输介质。通过直接与已建立的基于PCR的测试管道兼容的无RNA提取方案，可以显著节省时间和成本。这可能有助于扩大COVID-19测试。SARS冠状病毒2（SARS-CoV-2）感染的RT-PCR诊断方法被广泛采用，但RNA提取是快速、廉价诊断的瓶颈。在这里，作者开发了直接对热灭活的受试者样本或用现成的洗涤剂裂解的样本进行RT-PCR的方案，并对597份临床诊断的患者样本进行了基准性能测试。
Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is commonly diagnosed by reverse transcription polymerase chain reaction (RT-PCR) to detect viral RNA in patient samples, but RNA extraction constitutes a major bottleneck in current testing. Methodological simplification could increase diagnostic availability and efficiency, benefitting patient care and infection control. Here, we describe methods circumventing RNA extraction in COVID-19 testing by performing RT-PCR directly on heat-inactivated or lysed samples. Our data, including benchmarking using 597 clinical patient samples and a standardised diagnostic system, demonstrate that direct RT-PCR is viable option to extraction-based tests. Using controlled amounts of active SARS-CoV-2, we confirm effectiveness of heat inactivation by plaque assay and evaluate various generic buffers as transport medium for direct RT-PCR. Significant savings in time and cost are achieved through RNA-extraction-free protocols that are directly compatible with established PCR-based testing pipelines. This could aid expansion of COVID-19 testing. SARS-CoV-2 infection is widely diagnosed by RT-PCR, but RNA extraction is a bottleneck for fast and cheap diagnosis. Here, the authors develop protocols to perform RT-PCR directly on heat-inactivated subject samples or samples lysed with readily available detergents and benchmark performance against 597 clinically diagnosed patient samples.
登录
查看更多内容
影响因子：
19
作者：
Fomsgaard, Anna S.;Rosenstierne, Maiken Worsoe
通讯作者：
Rosenstierne, Maiken Worsoe
影响因子：
5.8
作者：
Kuhn, Max
通讯作者：
Kuhn, Max
DOI：
10.1016/j.jhin.2004.12.023
发表时间：
2005-10
期刊：
The Journal of hospital infection
影响因子：
--
作者：
Rabenau HF;Kampf G;Cinatl J;Doerr HW
通讯作者：
Doerr HW
影响因子：
4.9
作者：
Earl, Conner C.;Smith, Mark T.;Bundy, Bradley C.
通讯作者：
Bundy, Bradley C.
影响因子：
3.4
作者：
Khattari, Z;Brotons, G;Salditt, T
通讯作者：
Salditt, T