|Establishing a Puerto Rican Zika Virus stock||mmohns||2017-02-03|
Our lab has turned to using a ZIKV stock grown from a Puerto Rican isolate for our latest in vivo experiments (ZIKV-021, ZIKV-022, and ZIKV-023).ZIKV strain PRVABC59 (GenBank:KU501215), was originally isolated from a traveler to Puerto Rico. The seed stock passaged with three rounds of amplification on Vero cells was obtained from Brandy Russell (CDC, Ft. Collins, CO). Virus stocks were prepared by inoculation onto a confluent monolayer and passaged two times on C6/36 mosquito cells.Indian rhesus macaques challenged with 104 PFU PRVABC59 achieve peak viral load within the acute timeframe previously seen in inoculations with the French Polynesian ZIKV strain, albeit at a slightly lower peak (Figure 1a). Prolonged viremia has been detectable in pregnant animals, and is a consistent finding in an animal inoculated with PRVABC59 (Figure 1b).Additionally, 4 IFNAR-/- mice were inoculated in the left hind footpad with 106 PFU (in 50ul) PRVABC59. At 2 days post-infection, the mice were all successfully infected with viremias of 5.72, 5.51, 5.51, and 5.81 log10 PFU/ml (Figure 2). Mosquitoes were allowed to feed on the mice at this time point and then will be allowed to feed on macaques for a future transmission study.Sequencing data will be uploaded soon.