Lee, Dohwan Shin, Yong Chung, Seok Hwang, Kyo Seon Yoon, Dae Sung Lee, Jeong Hoon 2024-01-20T02:33:30Z 2024-01-20T02:33:30Z 2021-09-05 2016-12-20 0003-2700 https://pubs.kist.re.kr/handle/201004/123308 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. English AMER CHEMICAL SOC MEDIATED ISOTHERMAL AMPLIFICATION PCR INFECTION BRAZIL Simple and Highly Sensitive Molecular Diagnosis of Zika Virus by Lateral Flow Assays Article 10.1021/acs.analchem.6b03460 1 ANALYTICAL CHEMISTRY, v.88, no.24, pp.12272 - 12278 ANALYTICAL CHEMISTRY 88 24 12272 12278 scie scopus 000390621000043 2-s2.0-85028006246 Chemistry, Analytical Chemistry Article MEDIATED ISOTHERMAL AMPLIFICATION PCR INFECTION BRAZIL