A real-time reverse transcription polymerase chain reaction (RT-qPCR) assay that does not require Emergency Use Authorization (EUA) reagents was tested and validated for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the early stages of the outbreak of coronavirus disease 2019 (COVID-19) in the Republic of Korea. Early diagnosis of COVID-19 enables timely treatment and the implementation of public health measures. We validated the sensitivity, specificity, precision, linearity, accuracy, and robustness of the RT-qPCR assay for SARS-CoV-2 detection and compared its performance with that of several EUA-approved kits. Our RT-qPCR assay was highly specific for SARS-CoV-2 as demonstrated by not amplifying 13 other viruses that cause respiratory diseases. The assay showed high linearity using a viral isolate from a patient with known COVID-19 as well as plasmids containing target SARS-CoV-2 genes as templates. The assay showed good repeatability and reproducibility with a coefficient of variation of 3%, and a SARS-CoV-2 limit of detection of 1 PFU/mL. The RT-qPCR-based assay is highly effective and can facilitate the early diagnosis of COVID-19 without the use of EUA-approved kits or reagents in the Republic of Korea.
【저자키워드】 Virology, clinical microbiology, 【초록키워드】 COVID-19, Treatment, coronavirus disease, viruses, SARS-CoV-2, Coronavirus disease 2019, coronavirus, Variation, severe acute respiratory syndrome Coronavirus, respiratory diseases, EUA, early diagnosis, polymerase chain reaction, SARS-CoV-2 detection, sensitivity, specificity, RT-qPCR, Viral, Accuracy, outbreak, implementation, Patient, limit of detection, reverse transcription, respiratory, early stage, Emergency, Plasmid, Coefficient of variation, linearity, Chain Reaction, Real-Time Reverse Transcription Polymerase Chain Reaction, acute respiratory syndrome, Precision, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, reagents, public health measures, SARS-CoV-2 genes, early stages, coefficient, reproducibility, authorization, robustness, reagent, plasmids, effective, polymerase chain, virus, amplifying, facilitate, demonstrated, diagnosis of COVID-19, SARS-CoV-2 gene, was tested, 【제목키워드】 SARS-CoV-2, validation, COVID-19 outbreak, real-time RT-PCR, early stage,