Simultaneous detection of SARS-CoV-2 and identification of spike D614G mutation using point-of-care real-time polymerase chain reaction
- Authors
- Lee, So Yul; Lee, Ji Su; Ahn, Jeong Jin; Kim, Seung Jun; Sung, Heungsup; Huh, Jin Won; Hwang, Seung Yong
- Issue Date
- Jun-2022
- Publisher
- Elsevier BV
- Keywords
- SARS-CoV-2; Spike D614G mutation; Point-of-care; Real-time PCR; COVID-19; Molecular diagnosis
- Citation
- Journal of Virological Methods, v.304, pp.1 - 4
- Indexed
- SCIE
SCOPUS
- Journal Title
- Journal of Virological Methods
- Volume
- 304
- Start Page
- 1
- End Page
- 4
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/107690
- DOI
- 10.1016/j.jviromet.2022.114513
- ISSN
- 0166-0934
- 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. ~ 6 h); 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.
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