tetano
Editor, Senior Moderator
Arch Virol
. 2022 Feb 9.
doi: 10.1007/s00705-022-05383-0. Online ahead of print.
Diagnostic performance and clinical feasibility of a novel one-step RT-qPCR assay for simultaneous detection of multiple severe acute respiratory syndrome coronaviruses
Tran Bac Le[SUP] #[/SUP][SUP] 1 2 [/SUP], Hye Kwon Kim[SUP] #[/SUP][SUP] 3 [/SUP], Min-Ju Ahn[SUP] #[/SUP][SUP] 1 2 [/SUP], Mark Zanin[SUP] #[/SUP][SUP] 4 5 [/SUP], Van Thi Lo[SUP] 1 2 [/SUP], Shiman Ling[SUP] 4 [/SUP], Zhanpeng Jiang[SUP] 4 [/SUP], Jung-Ah Kang[SUP] 1 [/SUP], Pan Kee Bae[SUP] 6 [/SUP], Yeon-Sook Kim[SUP] 7 [/SUP], Seungtaek Kim[SUP] 8 [/SUP], Sook-San Wong[SUP] 9 10 [/SUP], Dae Gwin Jeong[SUP] 11 12 [/SUP], Sun-Woo Yoon[SUP] 13 14 [/SUP]
Affiliations
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° TCID[SUB]50[/SUB]/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.
. 2022 Feb 9.
doi: 10.1007/s00705-022-05383-0. Online ahead of print.
Diagnostic performance and clinical feasibility of a novel one-step RT-qPCR assay for simultaneous detection of multiple severe acute respiratory syndrome coronaviruses
Tran Bac Le[SUP] #[/SUP][SUP] 1 2 [/SUP], Hye Kwon Kim[SUP] #[/SUP][SUP] 3 [/SUP], Min-Ju Ahn[SUP] #[/SUP][SUP] 1 2 [/SUP], Mark Zanin[SUP] #[/SUP][SUP] 4 5 [/SUP], Van Thi Lo[SUP] 1 2 [/SUP], Shiman Ling[SUP] 4 [/SUP], Zhanpeng Jiang[SUP] 4 [/SUP], Jung-Ah Kang[SUP] 1 [/SUP], Pan Kee Bae[SUP] 6 [/SUP], Yeon-Sook Kim[SUP] 7 [/SUP], Seungtaek Kim[SUP] 8 [/SUP], Sook-San Wong[SUP] 9 10 [/SUP], Dae Gwin Jeong[SUP] 11 12 [/SUP], Sun-Woo Yoon[SUP] 13 14 [/SUP]
Affiliations
- PMID: 35137250
- DOI: 10.1007/s00705-022-05383-0
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° TCID[SUB]50[/SUB]/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.