title: An isothermal, label-free, and rapid one-step RNA amplification/detection assay for diagnosis of respiratory viral infections.
abstract: 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.
authors.affiliation: 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.
authors.forename: Bonhan
authors.initials: B
authors.lastname: Koo
authors.affiliation: 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.
authors.forename: Choong Eun
authors.initials: CE
authors.lastname: Jin
authors.affiliation: Department of Technology Education, Chungnam National University, Daejeon 34134, Republic of Korea.
authors.forename: Tae Yoon
authors.initials: TY
authors.lastname: Lee
authors.affiliation: Department of Electrical Engineering, Kwangwoon University, 447-1 Wolgye, Nowon, Seoul 136-791, Republic of Korea.
authors.forename: Jeong Hoon
authors.initials: JH
authors.lastname: Lee
authors.affiliation: One BioMed Pte Ltd, 60 Biopolis street, Genome #02-01, Singapore 138672, Singapore.
authors.forename: Mi Kyoung
authors.initials: MK
authors.lastname: Park
authors.affiliation: Department of Laboratory Medicine, Asan Medical Center, University of Ulsan College of Medicine, 88 Olympicro-43gil, Songpa-gu, Seoul, Republic of Korea.
authors.forename: Heungsup
authors.initials: H
authors.lastname: Sung
authors.affiliation: Department of Infectious Diseases, Asan Medical Center, University of Ulsan College of Medicine, 88 Olympicro-43gil, Songpa-gu, Seoul, Republic of Korea; Division of Infectious Diseases, Department of Internal Medicine, Soonchunhyang University College of Medicine, Seoul, Republic of Korea.
authors.forename: Se Yoon
authors.initials: SY
authors.lastname: Park
authors.affiliation: Department of Infectious Diseases, Asan Medical Center, University of Ulsan College of Medicine, 88 Olympicro-43gil, Songpa-gu, Seoul, Republic of Korea.
authors.forename: Hyun Jung
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Department of Infectious Diseases, Asan Medical Center, University of Ulsan College of Medicine, 88 Olympicro-43gil, Songpa-gu, Seoul, Republic of Korea.
authors.forename: Sun Mi
authors.initials: SM
authors.lastname: Kim
authors.affiliation: Department of Infectious Diseases, Asan Medical Center, University of Ulsan College of Medicine, 88 Olympicro-43gil, Songpa-gu, Seoul, Republic of Korea.
authors.forename: Ji Yeun
authors.initials: JY
authors.lastname: Kim
authors.affiliation: Department of Infectious Diseases, Asan Medical Center, University of Ulsan College of Medicine, 88 Olympicro-43gil, Songpa-gu, Seoul, Republic of Korea. Electronic address: kimsunghanmd@hotmail.com.
authors.forename: Sung-Han
authors.initials: SH
authors.lastname: Kim
authors.affiliation: 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. Electronic address: shinyongno1@gmail.com.
authors.forename: Yong
authors.initials: Y
authors.lastname: Shin
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D017934:Coronavirus / Q000235:genetics / Q000302:isolation & purification; D018352:Coronavirus Infections / Q000175:diagnosis / Q000235:genetics; D006801:Humans; D007251:Influenza, Human / Q000175:diagnosis / Q000235:genetics; D029521:Alphainfluenzavirus / Q000235:genetics / Q000302:isolation & purification; D029524:Betainfluenzavirus / Q000235:genetics / Q000302:isolation & purification; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D014777:Virus Diseases / Q000175:diagnosis* / Q000235:genetics
chemical_list: D012367:RNA, Viral
keywords: Emerging infectious disease; Label-free biosensor; RNA amplification/detection; Respiratory disease