SIV viral RNA (vRNA) levels in plasma were determined by real-time RT-PCR using the ABI Prism 7900 sequence detection system (Applied Biosystems) as previously described (79 (link)). Primer pairs for the assay corresponded to the forward nucleotides 1181 to 1208 and reverse nucleotides1338 to 1317 of the SIVmac239gag gene (80 (link)).
To assess ZIKV RNA levels in the plasma of our study RMs, viral RNA was extracted using EZ1 virus minikit v2.0 (Qiagen), and cDNA was synthesized with the SuperScript III first-strand synthesis system (Life Technologies). Absolute quantification of virus genome equivalents (GE; log10 per milliliter) was determined by real-time PCR using a TaqMan probe (Integrated DNA Technologies) targeting the ZIKV E protein. ZIKV RNA levels were measured in all animals to at least day 11 post-ZIKV infection. All animals, except RHDFiV (SIV/ZIKV cohort) measured below the limit of detection (LOD) of 3.2 log10 GE/ml at 11 days postinfection (dpi).
To assess ZIKV RNA levels in the plasma of our study RMs, viral RNA was extracted using EZ1 virus minikit v2.0 (Qiagen), and cDNA was synthesized with the SuperScript III first-strand synthesis system (Life Technologies). Absolute quantification of virus genome equivalents (GE; log10 per milliliter) was determined by real-time PCR using a TaqMan probe (Integrated DNA Technologies) targeting the ZIKV E protein. ZIKV RNA levels were measured in all animals to at least day 11 post-ZIKV infection. All animals, except RHDFiV (SIV/ZIKV cohort) measured below the limit of detection (LOD) of 3.2 log10 GE/ml at 11 days postinfection (dpi).
