Utilize este identificador para referenciar este registo:
https://hdl.handle.net/1822/75604
Título: | OmniSARS2: a highly sensitive and specific RT-qPCR-based COVID-19 diagnostic method designed to withstand SARS-CoV-2 lineage evolution |
Autor(es): | Carvalho-Correia, Eduarda Calçada, Carla Sofia Martins Branca, Fernando Estévez-Gómez, Nuria De Chiara, Loretta Varela, Nair Gallego-García, Pilar Posada, David Sousa, Hugo Sousa, João Carlos Veiga, Maria Isabel Osório, Nuno S. |
Palavras-chave: | SARS-CoV-2 COVID-19 RT-qPCR B.1.1.7 |
Data: | 26-Set-2021 |
Editora: | Multidisciplinary Digital Publishing Institute (MDPI) |
Revista: | Biomedicines |
Citação: | Carvalho-Correia, E.; Calçada, C.; Branca, F.; Estévez-Gómez, N.; De Chiara, L.; Varela, N.; Gallego-García, P.; Posada, D.; Sousa, H.; Sousa, J.; Veiga, M.I.; Osório, N.S. OmniSARS2: A Highly Sensitive and Specific RT-qPCR-Based COVID-19 Diagnostic Method Designed to Withstand SARS-CoV-2 Lineage Evolution. Biomedicines 2021, 9, 1314. https://doi.org/10.3390/biomedicines9101314 |
Resumo(s): | Extensive transmission of SARS-CoV-2 during the COVID-19 pandemic allowed the generation of thousands of mutations within its genome. While several of these become rare, others largely increase in prevalence, potentially jeopardizing the sensitivity of PCR-based diagnostics. Taking advantage of SARS-CoV-2 genomic knowledge, we designed a one-step probe-based multiplex RT-qPCR (OmniSARS2) to simultaneously detect short fragments of the SARS-CoV-2 genome in ORF1ab, E gene and S gene. Comparative genomics of the most common SARS-CoV-2 lineages, other human betacoronavirus and alphacoronavirus, was the basis for this design, targeting both highly conserved regions across SARS-CoV-2 lineages and variable or absent in other <i>Coronaviridae</i> viruses. The highest analytical sensitivity of this method for SARS-CoV-2 detection was 94.2 copies/mL at 95% detection probability (~1 copy per total reaction volume) for the S gene assay, matching the most sensitive available methods. In vitro specificity tests, performed using reference strains, showed no cross-reactivity with other human coronavirus or common pathogens. The method was compared with commercially available methods and detected the virus in clinical samples encompassing different SARS-CoV-2 lineages, including B.1, B.1.1, B.1.177 or B.1.1.7 and rarer lineages. OmniSARS2 revealed a sensitive and specific viral detection method that is less likely to be affected by lineage evolution oligonucleotide–sample mismatch, of relevance to ensure the accuracy of COVID-19 molecular diagnostic methods. |
Tipo: | Artigo |
URI: | https://hdl.handle.net/1822/75604 |
DOI: | 10.3390/biomedicines9101314 |
e-ISSN: | 2227-9059 |
Versão da editora: | https://www.mdpi.com/2227-9059/9/10/1314 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
Aparece nas coleções: |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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biomedicines-09-01314-v2.pdf | 1,94 MB | Adobe PDF | Ver/Abrir |
Este trabalho está licenciado sob uma Licença Creative Commons