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.
Keywords: COVID-19; Molecular diagnosis; Point-of-care; Real-time PCR; SARS-CoV-2; Spike D614G mutation.
【저자키워드】 COVID-19, SARS-CoV-2, point-of-care, Molecular diagnosis, Real-time PCR, Spike D614G mutation., 【초록키워드】 severe acute respiratory syndrome coronavirus 2, coronavirus, Mutation, spike, equipment, Human, Diagnosis, severe acute respiratory syndrome Coronavirus, Laboratory, respiratory viruses, polymerase chain reaction, point-of-care, specificity, rRT-PCR, PCR, D614G mutation, Sensitivity and specificity, Real-time PCR, POCT, D614G, multiplex, reverse transcription PCR, wild type, platform, Detection sensitivity, Analysis, strain, Chain Reaction, acute respiratory syndrome, rapid detection method, acute respiratory syndrome coronavirus, high mortality, probes, SARS-CoV-2 mutants, contagiousness, Multiple, labeling, probe, detection system, polymerase chain, detect, respiratory virus, applied, was reduced, SARS-CoV-2 mutant, 【제목키워드】 D614G mutation, Simultaneous, polymerase chain,