Table 1.
Generation and characterization of ZIKV sub-isolates used to derive PIZV candidates.
| Virus stock generation | Characterization | |
|---|---|---|
| CDC | Virus isolate amplified in Vero (2×) and C6/36 (1×)(V2/C6-1) | Plaque titer/phenotype, identity by RT-PCR, mycoplasma and togavirus testing |
| P1 | Virus amplification in Vero | TCID50 titer |
| P2 | Plaque purification of P1 | plaque phenotype |
| P3 | Plaque purification of P2 | plaque phenotype |
| P4 | Plaque purification of P3 | plaque phenotype |
| P5 | Amplification of P4 plaques (a-f sub-isolates) in Vero | TCID50 titer |
| P6 | Amplification of P5 virus (a-f sub-isolates) in Vero | TCID50 titer, plaque phenotype, full genome sequencing, growth kinetics in Vero |
| P7 | Amplification of P6 (b and e sub-isolates) in Vero; Used for deriving PIZV | Relative purity (SDS-PAGE), antigenicity (immunoblot, ELISA), EM, completeness of inactivation (serial cell culture passaging) |