Koo Bonhan, Jin Choong Eun, Lee Tae Yoon, Lee Jeong Hoon, Park Mi Kyoung, Sung Heungsup, Park Se Yoon, Lee Hyun Jung, Kim Sun Mi, Kim Ji Yeun, Kim Sung-Han, Shin Yong
Department of Convergence Medicine, Asan Medical Center, University of Ulsan College of Medicine, Biomedical Engineering Research Center, Asan Institute of Life Sciences, Asan Medical Center, 88 Olympicro-43gil, Songpa-gu, Seoul, Republic of Korea.
Department of Technology Education, Chungnam National University, Daejeon 34134, Republic of Korea.
Biosens Bioelectron. 2017 Apr 15;90:187-194. doi: 10.1016/j.bios.2016.11.051. Epub 2016 Nov 23.
Recently, RNA viral infections caused by respiratory viruses, such as influenza, parainfluenza, respiratory syncytial virus, coronavirus, and Middle East respiratory syndrome-coronavirus (MERS-CoV), and Zika virus, are a major public health threats in the world. Although myriads of diagnostic methods based on RNA amplification have been developed in the last decades, they continue to lack speed, sensitivity, and specificity for clinical use. A rapid and accurate diagnostic method is needed for appropriate control, including isolation and treatment of the patients. Here, we report an isothermal, label-free, one-step RNA amplification and detection system, termed as iROAD, for the diagnosis of respiratory diseases. It couples a one-step isothermal RNA amplification method and a bio-optical sensor for simultaneous viral RNA amplification/detection in a label-free and real-time manner. The iROAD assay offers a one-step viral RNA amplification/detection example to rapid analysis (<20min). The detection limit of iROAD assay was found to be 10-times more sensitive than that of real-time reverse transcription-PCR method. We confirmed the clinical utility of the iROAD assay by detecting viral RNAs obtained from 63 human respiratory samples. We envision that the iROAD assay will be useful and potentially adaptable for better diagnosis of emerging infectious diseases including respiratory diseases.
最近，由呼吸道病毒引起的RNA病毒感染，如流感病毒、副流感病毒、呼吸道合胞病毒、冠状病毒和中东呼吸综合征冠状病毒（MERS-CoV），以及寨卡病毒，是全球主要的公共卫生威胁。尽管在过去几十年中已经开发出无数基于RNA扩增的诊断方法，但它们在临床应用中仍然缺乏速度、灵敏度和特异性。需要一种快速准确的诊断方法来进行适当的控制，包括对患者的隔离和治疗。在此，我们报告一种用于诊断呼吸道疾病的等温、无标记、一步式RNA扩增和检测系统，称为iROAD。它将一步等温RNA扩增方法与生物光学传感器相结合，以无标记和实时方式同时进行病毒RNA的扩增/检测。iROAD检测提供了一个用于快速分析（<20分钟）的一步式病毒RNA扩增/检测实例。发现iROAD检测的检测限比实时逆转录PCR方法灵敏10倍。我们通过检测从63份人类呼吸道样本中获得的病毒RNA，证实了iROAD检测的临床实用性。我们设想，iROAD检测将有助于并可能适用于更好地诊断包括呼吸道疾病在内的新发传染病。
