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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
Title | Diagnostic performance and clinical feasibility of a novel one-step RT-qPCR assay for simultaneous detection of multiple severe acute respiratory syndrome coronaviruses |
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Authors | |
Issue Date | 2022 |
Citation | Archives of Virology, 2022, v. 167, n. 3, p. 871-879 How to Cite? |
Abstract | Coronavirus 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 Identifier | http://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 Field | Value | Language |
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dc.contributor.author | Le, Tran Bac | - |
dc.contributor.author | Kim, Hye Kwon | - |
dc.contributor.author | Ahn, Min Ju | - |
dc.contributor.author | Zanin, Mark | - |
dc.contributor.author | Lo, Van Thi | - |
dc.contributor.author | Ling, Shiman | - |
dc.contributor.author | Jiang, Zhanpeng | - |
dc.contributor.author | Kang, Jung Ah | - |
dc.contributor.author | Bae, Pan Kee | - |
dc.contributor.author | Kim, Yeon Sook | - |
dc.contributor.author | Kim, Seungtaek | - |
dc.contributor.author | Wong, Sook San | - |
dc.contributor.author | Jeong, Dae Gwin | - |
dc.contributor.author | Yoon, Sun Woo | - |
dc.date.accessioned | 2022-04-06T04:31:54Z | - |
dc.date.available | 2022-04-06T04:31:54Z | - |
dc.date.issued | 2022 | - |
dc.identifier.citation | Archives of Virology, 2022, v. 167, n. 3, p. 871-879 | - |
dc.identifier.issn | 0304-8608 | - |
dc.identifier.uri | http://hdl.handle.net/10722/311978 | - |
dc.description.abstract | Coronavirus 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.language | eng | - |
dc.relation.ispartof | Archives of Virology | - |
dc.rights | This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. | - |
dc.title | Diagnostic performance and clinical feasibility of a novel one-step RT-qPCR assay for simultaneous detection of multiple severe acute respiratory syndrome coronaviruses | - |
dc.type | Article | - |
dc.description.nature | published_or_final_version | - |
dc.identifier.doi | 10.1007/s00705-022-05383-0 | - |
dc.identifier.pmid | 35137250 | - |
dc.identifier.pmcid | PMC8885489 | - |
dc.identifier.scopus | eid_2-s2.0-85124349027 | - |
dc.identifier.volume | 167 | - |
dc.identifier.issue | 3 | - |
dc.identifier.spage | 871 | - |
dc.identifier.epage | 879 | - |
dc.identifier.eissn | 1432-8798 | - |
dc.identifier.isi | WOS:000752746400001 | - |