Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay.
Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay.
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DOI：
10.1371/journal.pone.0238612
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发表时间：
2020
期刊：
影响因子：
3.7
通讯作者：
Müller RU
中科院分区：
文献类型：
--
作者：
Schermer B;Fabretti F;Damagnez M;Di Cristanziano V;Heger E;Arjune S;Tanner NA;Imhof T;Koch M;Ladha A;Joung J;Gootenberg JS;Abudayyeh OO;Burst V;Zhang F;Klein F;Benzing T;Müller RU
对大部分人口和特定亚群进行快速和广泛的检测，对于妥善管理严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)感染和在大流行爆发时做出决策至关重要。然而，在急诊室、门诊、机场安检点或任何公共建筑的入口处进行护理点(POC)测试是一个重大挑战。热循环和核酸分离的需要阻碍了标准的基于聚合酶链式反应的方法用于这一目的。为了避免这些障碍，我们测试了从原始材料(鼻咽拭子)中检测SARS-CoV-2RNA的不依赖于PCR的方法，包括逆转录环介导的等温扩增(RT-LAMP)和特异性高灵敏度酶报告解锁(Sherlock)。虽然标准RT-LAMP检测的特异性似乎令人满意，但灵敏度尚未达到目前金标准的实时定量聚合酶链式反应(QPCR)检测。我们描述了一种新的多路RT-LAMP方法，并在原始样本上验证了其敏感性。这种方法可以快速、可靠地识别受感染的个人。引物的优化和多重有助于显著提高灵敏度。此外，我们还直接将我们的新型RT-LAMP检测方法与《神探夏洛克》进行了比较和结合。总而言之，这种方法揭示了一步多路RT-LAMP检测是开发简单且廉价的POC测试试剂盒的最佳选择。
Rapid and extensive testing of large parts of the population and specific subgroups is crucial for proper management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections and decision-making in times of a pandemic outbreak. However, point-of-care (POC) testing in places such as emergency units, outpatient clinics, airport security points or the entrance of any public building is a major challenge. The need for thermal cycling and nucleic acid isolation hampers the use of standard PCR-based methods for this purpose. To avoid these obstacles, we tested PCR-independent methods for the detection of SARS-CoV-2 RNA from primary material (nasopharyngeal swabs) including reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific high-sensitivity enzymatic reporter unlocking (SHERLOCK). Whilst specificity of standard RT-LAMP assays appears to be satisfactory, sensitivity does not reach the current gold-standard quantitative real-time polymerase chain reaction (qPCR) assays yet. We describe a novel multiplexed RT-LAMP approach and validate its sensitivity on primary samples. This approach allows for fast and reliable identification of infected individuals. Primer optimization and multiplexing helps to increase sensitivity significantly. In addition, we directly compare and combine our novel RT-LAMP assays with SHERLOCK. In summary, this approach reveals one-step multiplexed RT-LAMP assays as a prime-option for the development of easy and cheap POC test kits.
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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：
5.7
作者：
Huang, Wei E.;Lim, Boon;Cui, Zhanfeng
通讯作者：
Cui, Zhanfeng
通过逆转录环介导的等温扩增测定快速直观地检测 2019 新型冠状病毒 (SARS-CoV-2)
影响因子：
14.2
作者：
Yan, C.;Cui, J.;Yuan, J.
通讯作者：
Yuan, J.
影响因子：
5.8
作者：
Zhang, Rui;Liu, Jing;Zhou, Shao-Bing
通讯作者：
Zhou, Shao-Bing
影响因子：
11.8
作者：
Bullard, Jared;Dust, Kerry;Poliquin, Guillaume
通讯作者：
Poliquin, Guillaume