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Article: Diagnostic performance and clinical feasibility of a novel one-step RT-qPCR assay for simultaneous detection of multiple severe acute respiratory syndrome coronaviruses

TitleDiagnostic performance and clinical feasibility of a novel one-step RT-qPCR assay for simultaneous detection of multiple severe acute respiratory syndrome coronaviruses
Authors
Issue Date2022
Citation
Archives of Virology, 2022, v. 167, n. 3, p. 871-879 How to Cite?
AbstractCoronavirus disease 2019 (COVID-19) is an acute respiratory infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Other coronaviruses (CoVs) can also infect humans, although the majority cause only mild respiratory symptoms. Because early diagnosis of SARS-CoV-2 is critical for preventing further transmission events and improving clinical outcomes, it is important to be able to distinguish SARS-CoV-2 from other SARS-related CoVs in respiratory samples. Therefore, we developed and evaluated a novel reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay targeting the genes encoding the spike (S) and membrane (M) proteins to enable the rapid identification of SARS-CoV-2, including several new circulating variants and other emerging SARS-like CoVs. By analysis of in vitro-transcribed mRNA, we established multiplex RT-qPCR assays capable of detecting 5 × 10° copies/reaction. Using RNA extracted from cell culture supernatants, our multiple simultaneous SARS-CoV-2 assays had a limit of detection of 1 × 10° TCID50/mL and showed no cross-reaction with human CoVs or other respiratory viruses. We also validated our method using human clinical samples from patients with COVID-19 and healthy individuals, including nasal swab and sputum samples. This novel one-step multiplex RT-qPCR assay can be used to improve the laboratory diagnosis of human-pathogenic CoVs, including SARS-CoV-2, and may be useful for the identification of other SARS-like CoVs of zoonotic origin.
Persistent Identifierhttp://hdl.handle.net/10722/311978
ISSN
2023 Impact Factor: 2.5
2023 SCImago Journal Rankings: 0.590
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorLe, Tran Bac-
dc.contributor.authorKim, Hye Kwon-
dc.contributor.authorAhn, Min Ju-
dc.contributor.authorZanin, Mark-
dc.contributor.authorLo, Van Thi-
dc.contributor.authorLing, Shiman-
dc.contributor.authorJiang, Zhanpeng-
dc.contributor.authorKang, Jung Ah-
dc.contributor.authorBae, Pan Kee-
dc.contributor.authorKim, Yeon Sook-
dc.contributor.authorKim, Seungtaek-
dc.contributor.authorWong, Sook San-
dc.contributor.authorJeong, Dae Gwin-
dc.contributor.authorYoon, Sun Woo-
dc.date.accessioned2022-04-06T04:31:54Z-
dc.date.available2022-04-06T04:31:54Z-
dc.date.issued2022-
dc.identifier.citationArchives of Virology, 2022, v. 167, n. 3, p. 871-879-
dc.identifier.issn0304-8608-
dc.identifier.urihttp://hdl.handle.net/10722/311978-
dc.description.abstractCoronavirus disease 2019 (COVID-19) is an acute respiratory infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Other coronaviruses (CoVs) can also infect humans, although the majority cause only mild respiratory symptoms. Because early diagnosis of SARS-CoV-2 is critical for preventing further transmission events and improving clinical outcomes, it is important to be able to distinguish SARS-CoV-2 from other SARS-related CoVs in respiratory samples. Therefore, we developed and evaluated a novel reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay targeting the genes encoding the spike (S) and membrane (M) proteins to enable the rapid identification of SARS-CoV-2, including several new circulating variants and other emerging SARS-like CoVs. By analysis of in vitro-transcribed mRNA, we established multiplex RT-qPCR assays capable of detecting 5 × 10° copies/reaction. Using RNA extracted from cell culture supernatants, our multiple simultaneous SARS-CoV-2 assays had a limit of detection of 1 × 10° TCID50/mL and showed no cross-reaction with human CoVs or other respiratory viruses. We also validated our method using human clinical samples from patients with COVID-19 and healthy individuals, including nasal swab and sputum samples. This novel one-step multiplex RT-qPCR assay can be used to improve the laboratory diagnosis of human-pathogenic CoVs, including SARS-CoV-2, and may be useful for the identification of other SARS-like CoVs of zoonotic origin.-
dc.languageeng-
dc.relation.ispartofArchives of Virology-
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.-
dc.titleDiagnostic performance and clinical feasibility of a novel one-step RT-qPCR assay for simultaneous detection of multiple severe acute respiratory syndrome coronaviruses-
dc.typeArticle-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1007/s00705-022-05383-0-
dc.identifier.pmid35137250-
dc.identifier.pmcidPMC8885489-
dc.identifier.scopuseid_2-s2.0-85124349027-
dc.identifier.volume167-
dc.identifier.issue3-
dc.identifier.spage871-
dc.identifier.epage879-
dc.identifier.eissn1432-8798-
dc.identifier.isiWOS:000752746400001-

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