title: Simple and Highly Sensitive Molecular Diagnosis of Zika Virus by Lateral Flow Assays.
abstract: We have developed a simple, user-friendly, and highly sensitive Zika virus (ZIKV) detection method by incorporating optimized reverse transcription loop-mediated isothermal amplification (RT-LAMP) and a lateral flow assay (LFA). The optimized RT-LAMP reaction was carried out using Bst 3.0 polymerase, which has robust and fast isothermal amplification performance even in the presence of high concentrations of inhibitors; this permitted the amplification of ZIKV RNA in pure water and human whole blood. In addition, the strong reverse transcription activity of Bst 3.0 polymerase enabled specific ZIKV RNA amplification without extra addition of reverse transcriptase. The RT-LAMP condition was optimized by adjusting the Mg2+and dNTP mix concentration to extirpate nontarget amplification, which is caused by nonspecific primer dimers amplification. After 30 min of RT-LAMP reaction, the resultant amplicons were simply and rapidly analyzed by the LFA test in less than 5 min. The optimized RT-LAMP combined with the LFA allowed specific ZIKV RNA detection down to the single copy level within 35 min.
authors.affiliation: Department of Electrical Engineering, Kwangwoon University , 447-1 Wolgye, Nowon, Seoul 01897, Republic of Korea.
authors.forename: Dohwan
authors.initials: D
authors.lastname: Lee
authors.affiliation: Department of Convergence Medicine, Asan Medical Center, University of Ulsan College of Medicine , Ulsan, Republic of Korea.|Biomedical Engineering Research Center, Asan Institute of Life Sciences, Asan Medical Center , 88 Olympicro-43gil, Songpa-gu, Seoul, Republic of Korea.
authors.forename: Yong
authors.initials: Y
authors.lastname: Shin
authors.forename: Seok
authors.initials: S
authors.lastname: Chung
authors.affiliation: Center for BioMicrosystem, Korea Institute of Science and Technology (KIST) , Seoul 136-791, Korea.
authors.forename: Kyo Seon
authors.initials: KS
authors.lastname: Hwang
authors.forename: Dae Sung
authors.initials: DS
authors.lastname: Yoon
authors.affiliation: Department of Electrical Engineering, Kwangwoon University , 447-1 Wolgye, Nowon, Seoul 01897, Republic of Korea.
authors.forename: Jeong Hoon
authors.identifier: 0000-0001-6002-1557
authors.initials: JH
authors.lastname: Lee
mesh_terms: D004867:Equipment Design; D006801:Humans; D057230:Limit of Detection; D008274:Magnesium / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation* / Q000379:methods; D012367:RNA, Viral / Q000032:analysis* / Q000097:blood; D012680:Sensitivity and Specificity; D000071244:Zika Virus / Q000302:isolation & purification*; D000071243:Zika Virus Infection / Q000097:blood / Q000175:diagnosis* / Q000821:virology
chemical_list: D012367:RNA, Viral; D008274:Magnesium