Jee Hyunseul, Choi Minkyeong, Park In Su, Lee Junmin, Jang Woong Sik, Lim Chae Seung
BK21 Graduate Program, Department of Biomedical Sciences, College of Medicine, Korea University, Seoul 02841, Republic of Korea.
Emergency Medicine, College of Medicine, Korea University Guro Hospital, Seoul 08308, Republic of Korea.
Diagnostics (Basel). 2023 Sep 20;13(18):3001. doi: 10.3390/diagnostics13183001.
After three years of the SARS-CoV-2 pandemic, the demand for developing field-deployable point-of-care (PoC) molecular diagnostic tests has increased. Although RT-qPCR is the molecular diagnostic gold standard and is accurate, it is not readily applied to point-of-care testing (POCT). Meanwhile, rapid diagnostic kits have the disadvantage of low sensitivity. Recently, rapid isothermal nucleic acid amplification technology has emerged as an alternative for rapid diagnosis. Here, we developed a rapid SARS-CoV-2 reverse transcription loop-mediated isothermal amplification (RT-LAMP)-lateral flow assay (LFA) kit. This kit includes a Chelex-100/boiling nucleic acid extraction device and a one-step amplification detection apparatus capable of performing the entire process, from RNA extraction to detection, and diagnosing SARS-CoV-2 infection within 40 min without contamination. The detection limits of the rapid SARS-CoV-2 RT-LAMP-LFA kit were 100 plaque-forming units (PFUs) mL and 10 PFU mL for RNA samples extracted using the Chelex-100/boiling nucleic acid extraction device and commercial AdvansureTM E3 system, respectively. The sensitivity and specificity of the rapid SARS-CoV-2 RT-LAMP-LFA kit were 97.8% and 100%, respectively. Our SARS-CoV-2 RT-LAMP-LFA kit exhibited high sensitivity and specificity within 40 min without requiring laboratory instruments, suggesting that the kit could be used as a rapid POC molecular diagnostic test for SARS-CoV-2.
在新型冠状病毒肺炎大流行三年后，开发可现场部署的即时检测（PoC）分子诊断测试的需求有所增加。虽然逆转录定量聚合酶链反应（RT-qPCR）是分子诊断的金标准且准确，但它不易应用于即时检测（POCT）。同时，快速诊断试剂盒存在灵敏度低的缺点。最近，快速等温核酸扩增技术已成为快速诊断的替代方法。在此，我们开发了一种快速的新型冠状病毒2型逆转录环介导等温扩增（RT-LAMP）-侧向流动分析（LFA）试剂盒。该试剂盒包括一个Chelex-100/煮沸核酸提取装置和一个能够执行从RNA提取到检测的整个过程的一步式扩增检测装置，可在40分钟内诊断新型冠状病毒2型感染且无污染。使用Chelex-100/煮沸核酸提取装置和商用AdvansureTM E3系统提取的RNA样本，快速新型冠状病毒2型RT-LAMP-LFA试剂盒的检测限分别为100空斑形成单位（PFU）/mL和10 PFU/mL。快速新型冠状病毒2型RT-LAMP-LFA试剂盒的灵敏度和特异性分别为97.8%和100%。我们的新型冠状病毒2型RT-LAMP-LFA试剂盒在40分钟内显示出高灵敏度和特异性，无需实验室仪器，这表明该试剂盒可作为新型冠状病毒2型的快速即时检测分子诊断测试。
