Quantifying ZIKV RNA in Biological Fluids

Viral RNA levels in peripheral blood, urine, saliva, semen, CSF and cervicovaginal lavage (CVL) during primary infection (28 d after infection) were monitored by quantitative real-time PCR using primers and probe sets specific for ZIKV capsid. Viral RNA was isolated from cell-free blood plasma using QIAamp Viral RNA mini kit (Qiagen), as previously described^{32 (link)}. Viral RNA extraction from other samples were performed using the NucliSENS miniMAG System (BioMérieux) by following the manufacturer’s suggested instructions. Briefly, 500 μl of each sample was directly added to 2 ml of lysis buffer; a cervical vaginal swab was mixed in lysis buffer and eluted. Extracted RNA was used for amplification on a 7300 ABI Real-Time PCR system (Applied Biosystems).