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Suitcase Lab for Rapid Detection of SARS-CoV-2 Based on Recombinase Polymerase Amplification Assay

[Image: see text] In March 2020, the SARS-CoV-2 virus outbreak was declared as a world pandemic by the World Health Organization (WHO). The only measures for controlling the outbreak are testing and isolation of infected cases. Molecular real-time polymerase chain reaction (PCR) assays are very sens...

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Detalles Bibliográficos
Autores principales: El Wahed, Ahmed Abd, Patel, Pranav, Maier, Melanie, Pietsch, Corinna, Rüster, Dana, Böhlken-Fascher, Susanne, Kissenkötter, Jonas, Behrmann, Ole, Frimpong, Michael, Diagne, Moussa Moïse, Faye, Martin, Dia, Ndongo, Shalaby, Mohamed A., Amer, Haitham, Elgamal, Mahmoud, Zaki, Ali, Ismail, Ghada, Kaiser, Marco, Corman, Victor M., Niedrig, Matthias, Landt, Olfert, Faye, Ousmane, Sall, Amadou A., Hufert, Frank T., Truyen, Uwe, Liebert, Uwe G., Weidmann, Manfred
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7839158/
https://www.ncbi.nlm.nih.gov/pubmed/33471510
http://dx.doi.org/10.1021/acs.analchem.0c04779
Descripción
Sumario:[Image: see text] In March 2020, the SARS-CoV-2 virus outbreak was declared as a world pandemic by the World Health Organization (WHO). The only measures for controlling the outbreak are testing and isolation of infected cases. Molecular real-time polymerase chain reaction (PCR) assays are very sensitive but require highly equipped laboratories and well-trained personnel. In this study, a rapid point-of-need detection method was developed to detect the RNA-dependent RNA polymerase (RdRP), envelope protein (E), and nucleocapsid protein (N) genes of SARS-CoV-2 based on the reverse transcription recombinase polymerase amplification (RT-RPA) assay. RdRP, E, and N RT-RPA assays required approximately 15 min to amplify 2, 15, and 15 RNA molecules of molecular standard/reaction, respectively. RdRP and E RT-RPA assays detected SARS-CoV-1 and 2 genomic RNA, whereas the N RT-RPA assay identified only SARS-CoV-2 RNA. All established assays did not cross-react with nucleic acids of other respiratory pathogens. The RT-RPA assay’s clinical sensitivity and specificity in comparison to real-time RT-PCR (n = 36) were 94 and 100% for RdRP; 65 and 77% for E; and 83 and 94% for the N RT-RPA assay. The assays were deployed to the field, where the RdRP RT-RPA assays confirmed to produce the most accurate results in three different laboratories in Africa (n = 89). The RPA assays were run in a mobile suitcase laboratory to facilitate the deployment at point of need. The assays can contribute to speed up the control measures as well as assist in the detection of COVID-19 cases in low-resource settings.