Fig. 3.

Cross-template detection of the different ZIKV strain targets using the ZIKV RT-RPA. (a) Detection of different ZIKV synthetic RNA templates (1–5) using the ZIKV RT-RPA assay. All synthetic fragments were used at 5×10³ copies per reaction and compared to the non-template control (NTC). (b) Amplification of target region in the ZIKV RT-RPA assay using extracted nucleic acid from two strains of cultured ZIKV (African – KY288905 and South American – KU501215) compared to the detection of Zika synthetic RNA fragment 5. Both cultured viral strains were used in the assay at 1.5×10¹ p.f.u. per reaction, whereas the synthetic fragment 5 was used at 5×10³ copies per reaction. The fluorescent signal generated by the non-template control (NTC) is also shown for reference. Amplification curves show the average total fluorescence values from three independent ZIKV RT-RPA assays; standard deviations are represented as error bars.