Zika virus lateral flow assays using reverse transcription-loop-mediated isothermal amplification. Issue 29 (18th May 2021)

• Record Type: Journal Article
• Authors: Ahn, Gna; Lee, SeonHyung; Lee, Se Hee; Baek, Yun Hee; Song, Min-Suk; Kim, Yang-Hoon; Ahn, Ji-Young
• Is Part Of: RSC advances. Volume 11:Issue 29(2021)
• Page Start: 17800
• Page End: 17808
• Publication Date: 2021-05-18
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/d1ra01227d
• Languages: English
• ISSNs: 2046-2069

Abstract

Abstract : Our study suggest that ZIKV RT-LAMP combined with LFA could serve as a rapid, accurate, and independent point-of-care detection method for ZIKV outbreaks. Abstract : Owing to the global spread of the Zika virus (ZIKV) infection, field-ready diagnostics are urgently warranted. In this study, we sought to detect ZIKV using reverse transcription loop-mediated isothermal amplification (RT-LAMP). Briefly, we performed and optimized ZIKV RT-LAMP for the analysis of biological samples (PBS, urine, and plasma). Based on our findings, this method could detect ZIKV RNA in 40 min at 63 °C without any off-target amplification. After performing specificity tests using BtsI restriction enzyme digestion, the feasibility of ZIKV RT-LAMP was determined via end-point detection with different sample matrices. Thereafter, a lateral flow assay (LFA) was conducted to directly detect the ZIKV RT-LAMP products. Based on the LFA reaction, hybridization occurred between the AuNPs:polyadenylated (polyA10 )-ZIKV probe and the LAMP amplicons. Subsequently, we optimized the assay parameters, including the concentration of AuNPs and migration matrices (glass fiber and nitrocellulose membrane). By employing a specific AuNP:polyA10 -ZIKV LAMP probe, we could demonstrate the purpose and utility of primary and secondary antibodies. Owing to LFA, the resultant ZIKV RT-LAMP products were rapidly and simply assayed in less than 5 min. Further, no preparation step was required to achieve LAMP-probe hybridization, highlighting the utility of this method for field-ready ZIKV diagnosis. Collectively, our findings suggest that ZIKV RT-LAMP combined with LFA could serve as a rapid, accurate, and independent point-of-care detection method for preventing ZIKV outbreaks.