Rapid detection of novel coronavirus/Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) by reverse transcription-loop-mediated isothermal amplification
Rapid detection of novel coronavirus/Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) by reverse transcription-loop-mediated isothermal amplification
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DOI：
10.1371/journal.pone.0234682
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发表时间：
2020-06-12
期刊：
影响因子：
3.7
通讯作者：
Chancellor, Michael B.
中科院分区：
文献类型：
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作者：
Lamb, Laura E.;Bartolone, Sarah N.;Chancellor, Michael B.
新型冠状病毒/严重急性呼吸道综合征冠状病毒2（SARS-CoV-2或2019-nCoV）以及由该病毒引起的后续疾病（2019冠状病毒病或COVID-19）是一种新兴的全球健康问题，需要快速诊断测试。定量逆转录PCR（qRT-PCR）是目前SARS-CoV-2检测的标准;然而，逆转录环介导的等温扩增（RT-LAMP）可能允许在风险点进行更快，更便宜的现场检测。这项研究的目的是开发一种快速筛查诊断测试，可以在30-45分钟内完成。通过用SARS-CoV-2核酸序列的一部分掺入血清、尿液、唾液、口咽拭子和鼻咽拭子来产生模拟患者样本。还测试了从实际COVID-19患者收集的鼻咽拭子中分离的RNA。使用RT-LAMP以及通过常规qRT-PCR测试样品。RT-LAMP的特异性也通过针对其他相关冠状病毒的测试来评估。RT-LAMP在模拟患者样本和临床标本中均特异性检测到SARS-CoV-2。该测试在30-45分钟内进行。这种方法可用于监测受影响的个人，或可能有助于现场和潜在入境口岸的筛查工作。
Novel Corona virus/Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2 or 2019-nCoV), and the subsequent disease caused by the virus (coronavirus disease 2019 or COVID-19), is an emerging global health concern that requires a rapid diagnostic test. Quantitative reverse transcription PCR (qRT-PCR) is currently the standard for SARS-CoV-2 detection; however, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for faster and cheaper field based testing at point-of-risk. The objective of this study was to develop a rapid screening diagnostic test that could be completed in 30-45 minutes. Simulated patient samples were generated by spiking serum, urine, saliva, oropharyngeal swabs, and nasopharyngeal swabs with a portion of the SARS-CoV-2 nucleic sequence. RNA isolated from nasopharyngeal swabs collected from actual COVID-19 patients was also tested. The samples were tested using RT-LAMP as well as by conventional qRT-PCR. Specificity of the RT-LAMP was evaluated by also testing against other related coronaviruses. RT-LAMP specifically detected SARS-CoV-2 in both simulated patient samples and clinical specimens. This test was performed in 30-45 minutes. This approach could be used for monitoring of exposed individuals or potentially aid with screening efforts in the field and potential ports of entry.