Chung Yoon-Seok, Lee Nam-Joo, Woo Sang Hee, Kim Jeong-Min, Kim Heui Man, Jo Hye Jun, Park Ye Eun, Han Myung-Guk
Division of Infectious Disease Diagnosis Control, Honam Regional Centers for Disease Control and Prevention, Korea Diseases Control and Prevention Agency, Gwangju-si, 61947, Republic of Korea.
Division of Emerging Infectious Diseases, Bureau of Infectious Disease Diagnosis Control, Korea Diseases Control and Prevention Agency, Cheongju-si, 28159, Republic of Korea.
Sci Rep. 2021 Jul 20;11(1):14817. doi: 10.1038/s41598-021-94196-3.
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.
一种无需紧急使用授权（EUA）试剂的实时逆转录聚合酶链反应（RT-qPCR）检测方法，在韩国2019冠状病毒病（COVID-19）疫情爆发初期，针对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的检测进行了测试和验证。COVID-19的早期诊断有助于及时治疗和实施公共卫生措施。我们验证了用于SARS-CoV-2检测的RT-qPCR检测方法的灵敏度、特异性、精密度、线性、准确性和稳健性，并将其性能与几种经EUA批准的试剂盒进行了比较。我们的RT-qPCR检测方法对SARS-CoV-2具有高度特异性，未扩增其他13种引起呼吸道疾病的病毒即可证明。该检测方法以已知COVID-19患者的病毒分离株以及含有目标SARS-CoV-2基因的质粒为模板，显示出高线性。该检测方法具有良好的重复性和再现性，变异系数为3%，SARS-CoV-2检测限为1 PFU/mL。基于RT-qPCR的检测方法非常有效，在韩国无需使用经EUA批准的试剂盒或试剂即可促进COVID-19的早期诊断。
