tetano
Editor, Senior Moderator
J Virol Methods
. 2022 Mar 4;114513.
doi: 10.1016/j.jviromet.2022.114513. Online ahead of print.
Simultaneous detection of SARS-CoV-2 and identification of spike D614G mutation using point-of-care real-time polymerase chain reaction
So Yul Lee[SUP] 1 [/SUP], Ji Su Lee[SUP] 1 [/SUP], Jeong Jin Ahn[SUP] 2 [/SUP], Seung Jun Kim[SUP] 2 [/SUP], Heungsup Sung[SUP] 3 [/SUP], Jin Won Huh[SUP] 4 [/SUP], Seung Yong Hwang[SUP] 5 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is associated with high mortality and infectivity rates in humans since its emergence. Analysis using high-accuracy real-time polymerase chain reaction (PCR) is recommended for the detection of general respiratory viruses including SARS-CoV-2, but it takes a long time (e.g. ~ 6hr); moreover, on-site diagnosis is difficult owing to the need for skilled technicians and advanced laboratory facilities. Currently, the importance of point-of-care testing (POCT) is being emphasized for the rapid detection of SARS-CoV-2. Here, we developed a multiplex real-time reverse transcription PCR (rRT-PCR) analysis that not only detects SARS-CoV-2 but also D614G strains with higher contagiousness than wild types among SARS-CoV-2 mutants using probe-based rRT-PCR. Moreover, this method was applied to portable PCR equipment capable of POCT to confirm high detection sensitivity and specificity. Multiple assays were possible with fluorescence labeling of individual probes. Furthermore, using a microfluidic chip-based point-of-care testing rRT-PCR platform, detection time was reduced by more than half compared with the commonly used detection system. This demonstrates that our assay has 100% of high sensitivity and specificity and could thus aid in the rapid and simple screening of SARS-CoV-2 carrying the mutation. We present a rapid detection method for mutations in SARS-CoV-2.
Keywords: COVID-19; SARS-CoV-2; molecular diagnosis; point-of-care; real-time PCR; spike D614G mutation.
. 2022 Mar 4;114513.
doi: 10.1016/j.jviromet.2022.114513. Online ahead of print.
Simultaneous detection of SARS-CoV-2 and identification of spike D614G mutation using point-of-care real-time polymerase chain reaction
So Yul Lee[SUP] 1 [/SUP], Ji Su Lee[SUP] 1 [/SUP], Jeong Jin Ahn[SUP] 2 [/SUP], Seung Jun Kim[SUP] 2 [/SUP], Heungsup Sung[SUP] 3 [/SUP], Jin Won Huh[SUP] 4 [/SUP], Seung Yong Hwang[SUP] 5 [/SUP]
Affiliations
- PMID: 35257681
- DOI: 10.1016/j.jviromet.2022.114513
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is associated with high mortality and infectivity rates in humans since its emergence. Analysis using high-accuracy real-time polymerase chain reaction (PCR) is recommended for the detection of general respiratory viruses including SARS-CoV-2, but it takes a long time (e.g. ~ 6hr); moreover, on-site diagnosis is difficult owing to the need for skilled technicians and advanced laboratory facilities. Currently, the importance of point-of-care testing (POCT) is being emphasized for the rapid detection of SARS-CoV-2. Here, we developed a multiplex real-time reverse transcription PCR (rRT-PCR) analysis that not only detects SARS-CoV-2 but also D614G strains with higher contagiousness than wild types among SARS-CoV-2 mutants using probe-based rRT-PCR. Moreover, this method was applied to portable PCR equipment capable of POCT to confirm high detection sensitivity and specificity. Multiple assays were possible with fluorescence labeling of individual probes. Furthermore, using a microfluidic chip-based point-of-care testing rRT-PCR platform, detection time was reduced by more than half compared with the commonly used detection system. This demonstrates that our assay has 100% of high sensitivity and specificity and could thus aid in the rapid and simple screening of SARS-CoV-2 carrying the mutation. We present a rapid detection method for mutations in SARS-CoV-2.
Keywords: COVID-19; SARS-CoV-2; molecular diagnosis; point-of-care; real-time PCR; spike D614G mutation.