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dc.contributor.authorJones, Les
dc.contributor.authorBakre, Abhijeet
dc.contributor.authorNaikare, Hemant
dc.contributor.authorKolhe, Ravindra
dc.contributor.authorSanchez, Susan
dc.contributor.authorMosley, Yung-Yi C.
dc.contributor.authorTripp, Ralph A.
dc.date.accessioned2022-03-09T17:41:34Z
dc.date.available2022-03-09T17:41:34Z
dc.date.issued2021-09-17
dc.identifier.doi10.1371/journal.pone.0257563
dc.identifier.urihttp://hdl.handle.net/10675.2/624243
dc.description.abstractThe 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.sponsorshipFunding was through the Georgia COVID19 Task Force and the Georgia Research Alliance to RT.en_US
dc.language.isoenen_US
dc.publisherPublic Library of Science (PLoS)en_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectMultidisciplinaryen_US
dc.titleIsothermal amplification and fluorescent detection of SARS-CoV-2 and SARS-CoV-2 variant virus in nasopharyngeal swabsen_US
dc.typeArticleen_US
dc.identifier.eissn1932-6203
dc.contributor.departmentDepartment of Pathologyen_US
dc.identifier.journalPloSen_US
dc.source.journaltitlePLOS ONE
dc.source.volume16
dc.source.issue9
dc.source.beginpagee0257563
refterms.dateFOA2022-03-09T17:41:34Z


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