Park Gun-Soo, Baek Seung-Hwa, Ku Keunbon, Kim Seong-Jun, Kim Seung Il, Kim Bum-Tae, Maeng Jin-Soo
Center for Convergent Research of Emerging Virus Infection, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea.
Research Group of Food Processing, Korea Food Research Institute, Wanju-gun, Jeollabuk-do, Republic of Korea.
Bio Protoc. 2020 Nov 5;10(21):e3804. doi: 10.21769/BioProtoc.3804.
Standard diagnostic methods of Coronavirus Disease 2019 (COVID-19) rely on RT-qPCR technique which have limited point-of-care test (POCT) potential due to necessity of dedicated equipment and specialized personnel. LAMP, an isothermal nucleic acid amplification test (NAAT), is a promising technique that may substitute RT-qPCR for POCT of genomic materials. Here, we provide a protocol to perform reverse transcription LAMP targeting SARS-CoV-2. We adopted both real-time fluorescence detection and end-point colorimetric detection approaches. Our protocol would be useful for screening diagnosis of COVID-19 and be a baseline for development of improved POCT NAAT.
2019年冠状病毒病(COVID-19)的标准诊断方法依赖于逆转录定量聚合酶链反应(RT-qPCR)技术,由于需要专用设备和专业人员,其即时检测(POCT)潜力有限。环介导等温扩增技术(LAMP)是一种等温核酸扩增检测(NAAT),是一种很有前景的技术,可替代RT-qPCR用于基因组材料的POCT。在此,我们提供了一种针对严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)进行逆转录LAMP的方案。我们采用了实时荧光检测和终点比色检测方法。我们的方案将有助于COVID-19的筛查诊断,并为改进POCT NAAT的开发提供基线。