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Rapid Detection of SARS-CoV-2 RNA Using Reverse Transcription Recombinase Polymerase Amplification (RT-RPA) with Lateral Flow for N-Protein Gene and Variant-Specific Deletion-Insertion Mutation in S-Protein Gene.

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dc.contributor.author Malaga, Jose L.
dc.contributor.author Pajuelo Travezaño, Monica Jhenny
dc.contributor.author Okamoto, Michiko
dc.contributor.author Tsinda, Emmanuel Kagning
dc.contributor.author Otani, Kanako
dc.contributor.author Tsukayama Cisneros, Pablo
dc.contributor.author Mascaro Rivera, Lucero Nancy
dc.contributor.author Cuicapuza Artega, Diego Bernhard
dc.contributor.author Katsumi, Masamichi
dc.contributor.author Kawamura, Kazuhisa
dc.contributor.author Nishimura, Hidekazu
dc.contributor.author Sakagami, Akie
dc.contributor.author Ueki, Yo
dc.contributor.author Omiya, Suguru
dc.contributor.author Okamoto, Satoshi
dc.contributor.author Nakayama, Asami
dc.contributor.author Fujimaki, Shin-Ichi
dc.contributor.author Yu, Chuyao
dc.contributor.author Azam, Sikandar
dc.contributor.author Kodama, Eiichi
dc.contributor.author Dapat, Clyde
dc.contributor.author Oshitani, Hitoshi
dc.contributor.author Saito, Mayuko
dc.date.accessioned 2023-07-18T16:18:56Z
dc.date.available 2023-07-18T16:18:56Z
dc.date.issued 2023
dc.identifier.uri https://hdl.handle.net/20.500.12866/13929
dc.description.abstract Rapid molecular testing for severe acute respiratory coronavirus 2 (SARS-CoV-2) variants may contribute to the development of public health measures, particularly in resource-limited areas. Reverse transcription recombinase polymerase amplification using a lateral flow assay (RT-RPA-LF) allows rapid RNA detection without thermal cyclers. In this study, we developed two assays to detect SARS-CoV-2 nucleocapsid (N) gene and Omicron BA.1 spike (S) gene-specific deletion-insertion mutations (del211/ins214). Both tests had a detection limit of 10 copies/µL in vitro and the detection time was approximately 35 min from incubation to detection. The sensitivities of SARS-CoV-2 (N) RT-RPA-LF by viral load categories were 100% for clinical samples with high (>9015.7 copies/µL, cycle quantification (Cq): < 25) and moderate (385.5-9015.7 copies/µL, Cq: 25-29.9) viral load, 83.3% for low (16.5-385.5 copies/µL, Cq: 30-34.9), and 14.3% for very low (<16.5 copies/µL, Cq: 35-40). The sensitivities of the Omicron BA.1 (S) RT-RPA-LF were 94.9%, 78%, 23.8%, and 0%, respectively, and the specificity against non-BA.1 SARS-CoV-2-positive samples was 96%. The assays seemed more sensitive than rapid antigen detection in moderate viral load samples. Although implementation in resource-limited settings requires additional improvements, deletion-insertion mutations were successfully detected by the RT-RPA-LF technique. en_US
dc.language.iso eng
dc.publisher MDPI
dc.relation.ispartofseries Viruses
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject SARS-CoV-2 en_US
dc.subject Variant of concern (VOC) en_US
dc.subject Deletion–insertion mutation en_US
dc.subject COVID-19 en_US
dc.subject Recombinase polymerase amplification (RPA) en_US
dc.subject.mesh SARS-CoV-2
dc.subject.mesh Mutación
dc.subject.mesh COVID-19
dc.subject.mesh Reacción en Cadena de la Polimerasa
dc.title Rapid Detection of SARS-CoV-2 RNA Using Reverse Transcription Recombinase Polymerase Amplification (RT-RPA) with Lateral Flow for N-Protein Gene and Variant-Specific Deletion-Insertion Mutation in S-Protein Gene. en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.3390/v15061254
dc.relation.issn 1999-4915


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