Table 2.
Summary of the experiments demonstrating the replication of the recovered viruses.
| Viral strain (Accession Number) | Template for subgenomic amplicons | Cell line used | CPE | Amount of viral RNA (Log 10 copies/mL; mean + /− SD) | infectious titers(Log 10 TCID50/mL; mean + /− SD) | Complete genome similarity | ||
|---|---|---|---|---|---|---|---|---|
| Origin | Number of fragments | DpnI treatment | Transfection/ Passage | |||||
| JEV (KC196115) | DNS | 3 | No | U4.4 | Yes | 6.82+/− 0.20 | 6.40+/− 0.10 | 99.9% |
| DNS | 3 | No | C6/36 | Yes | 7.22+/− 0.21 | 8.41+/− 0.29 | 99.9% | |
| DNS | 3 | No | BHK-21 | Yes | 7.09+/− 0.18 | 7.69+/− 0.12 | 99.9% | |
| DNS | 3 | No | HEK-293 | Yes | 6,91+/− 0.34 | 7.98+/− 0.24 | 99.9% | |
| DNS | 3 | No | SW13 | Yes | 6.65+/− 0.23 | 7.65+/− 0.24 | 99.9% | |
| DNS | 4 | Yes | C6/36 | Yes | 7.34+/− 0.09 | 7.88+/− 0.43 | 99.9% | |
| DNS | 5 | Yes | C6/36 | Yes | 7.28+/− 0.04 | 7.46+/− 0.10 | 99.9% | |
| DNS | 6 | Yes | C6/36 | Yes | 7.33+/− 0.07 | 7.69+/− 0.12 | 99.9% | |
| DNS | 8 | Yes | C6/36 | Yes | 6.95+/− 0.14 | 7.37+/− 0.28 | 99.9% | |
| DNS | 10 | Yes | C6/36 | Yes | 6.66+/− 0.06 | 6.49+/− 0.07 | 99.9% | |
| ChikV (EU224268) | I.C. | 3 | Yes | C6/36 | No | 7.44+/− 0.09 | 5.65+/− 0.12 | NA |
| YFV Asibi (AY640589) | DNS | 3 | No | C6/36 | Yes | 7.18+/− 0.10 | 7.92+/− 0.2 | NA |
| DNS | 3 | No | HEK-293 | Yes | 7.15+/− 0.08 | 6.26+/− 0.38 | NA | |
| DNS | 3 | No | BHK-21 | Yes | 7.13+/− 0.04 | 6.34+/− 0.18 | NA | |
| YFV 17D (NC002031) | DNS | 3 | No | C6/36 | No | 5.41+/− 0,26 | 6.56+/− 0.17 | NA |
| DNS | 3 | No | HEK-293 | Yes | 6.39+/− 0.04 | 6.22+/− 0.13 | NA | |
| DNS | 3 | No | BHK-21 | Yes | 6.24+/− 0.08 | 6.48+/− 0.05 | NA | |
| Zika PF (KJ776791) | DNS | 3 | No | C6/36 | Yes | 8.38+/− 0.21 | 4.04+/− 0.14 | NA |
| Zika DAK (KU955592) | DNS | 3 | No | C6/36 | Yes | 8.58+/− 0.38 | 5.14+/− 0.07 | NA |
Description of the origin of the initial material used as template for the production of the overlapping PCR fragments (DNS, de novo Synthesis; I.C., infectious clone), the number of transfected amplicons and their treatment by the DpnI restriction enzyme before transfection, the cell lines used for transfection and passages, the presence or absence of cytopathic effect (CPE), the amounts of viral RNA in cell supernatant at the fourth passage by real-time RT-PCR, the infectious titers in cell supernatant at the second passage by TCID50 assay and the complete genome similarity assay. For amounts of viral RNA and infectious titers, mean values +/− SD values were represented. *NA: not available.