Fig. 3.

Real-time RT-LAMP on Zika detection (ZV). a Singleplex and multiplexed LOD for Zika using 80 nM FAM-labeled strand-displacing probe. Zika viral titers ranged from 2.85 pfu to 0.71 pfu. Fluorescent emission upon irradiation by a blue LED (470 nm) was visualized through orange filter glass after incubation at 65 °C for 30 min. b Cross-reactivity assay for Zika where fluorescent probes for chikungunya (HEX) and dengue-1 (TAMRA) were excluded and only FAM-labeled amplicons could be detected. Viral titers were as follows: 2.85 pfu for ZV, 242 genomic copies for CH, and 1.22 pfu for D-1. Visualization was done as before after incubation at 65 °C for 35 min. c Tolerance to urine in RT-LAMP. Viral titer of Zika RNA was 2.85 pfu. Varying concentrations (0%-50%) of urine were tested using 80 nM FAM-labeled strand-displacing probe