*2.6. ZIKV Viral Stock Productuion*

A primary clinical virus specimen was isolated from a serum sample of a patient from Paraiba. The virus was propagated in a culture of C6/36 *Ae. albopictus* mosquito cells and harvested virus was tittered by the infection of Vero cells (CCL-81) followed by RT-PCRq, which determined a titer of 5.8 × 10<sup>6</sup> PFU/mL. Copy numbers were assessed by using a standard curve in the RT-PCRq reaction containing 1 × 10<sup>8</sup> copies/reaction. The oligonucleotide set utilized targeted the intergenic region of the Membrane/Envelope as described by Lanciotti, 2008 [26] (Table 1).


**Table 1.** Oligonucleotide sets to amplify ZIKV genome.
