Isothermal amplification and fluorescent detection of SARS-CoV-2 and SARS-CoV-2 variant virus in nasopharyngeal swabs
dc.contributor.author | Jones, Les | |
dc.contributor.author | Bakre, Abhijeet | |
dc.contributor.author | Naikare, Hemant | |
dc.contributor.author | Kolhe, Ravindra | |
dc.contributor.author | Sanchez, Susan | |
dc.contributor.author | Mosley, Yung-Yi C. | |
dc.contributor.author | Tripp, Ralph A. | |
dc.date.accessioned | 2022-03-09T17:41:34Z | |
dc.date.available | 2022-03-09T17:41:34Z | |
dc.date.issued | 2021-09-17 | |
dc.identifier.doi | 10.1371/journal.pone.0257563 | |
dc.identifier.uri | http://hdl.handle.net/10675.2/624243 | |
dc.description.abstract | The COVID-19 pandemic caused by the SARS-CoV-2 is a serious health threat causing worldwide morbidity and mortality. Real-time reverse transcription PCR (RT-qPCR) is currently the standard for SARS-CoV-2 detection. Although various nucleic acid-based assays have been developed to aid the detection of SARS-CoV-2 from COVID-19 patient samples, the objective of this study was to develop a diagnostic test that can be completed in 30 minutes without having to isolate RNA from the samples. Here, we present an RNA amplification detection method performed using reverse transcription loop-mediated isothermal amplification (RT-LAMP) reactions to achieve specific, rapid (30 min), and sensitive (<100 copies) fluorescent detection in real-time of SARS-CoV-2 directly from patient nasopharyngeal swab (NP) samples. When compared to RT-qPCR, positive NP swab samples assayed by fluorescent RT-LAMP had 98% (n = 41/42) concordance and negative NP swab samples assayed by fluorescent RT-LAMP had 87% (n = 59/68) concordance for the same samples. Importantly, the fluorescent RT-LAMP results were obtained without purification of RNA from the NP swab samples in contrast to RT-qPCR. We also show that the fluorescent RTLAMP assay can specifically detect live virus directly from cultures of both SARS-CoV-2 wild type (WA1/2020), and a SARS-CoV-2 B.1.1.7 (alpha) variant strain with equal sensitivity to RT-qPCR. RT-LAMP has several advantages over RT-qPCR including isothermal amplification, speed (<30 min), reduced costs, and similar sensitivity and specificity. | en_US |
dc.description.sponsorship | Funding was through the Georgia COVID19 Task Force and the Georgia Research Alliance to RT. | en_US |
dc.language.iso | en | en_US |
dc.publisher | Public Library of Science (PLoS) | en_US |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.subject | Multidisciplinary | en_US |
dc.title | Isothermal amplification and fluorescent detection of SARS-CoV-2 and SARS-CoV-2 variant virus in nasopharyngeal swabs | en_US |
dc.type | Article | en_US |
dc.identifier.eissn | 1932-6203 | |
dc.contributor.department | Department of Pathology | en_US |
dc.identifier.journal | PloS | en_US |
dc.source.journaltitle | PLOS ONE | |
dc.source.volume | 16 | |
dc.source.issue | 9 | |
dc.source.beginpage | e0257563 | |
refterms.dateFOA | 2022-03-09T17:41:34Z |
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COVID-19 Publications [97]
Publications by University Affiliates Related to COVID-19