INTRODUCTION: Zika virus is mainly transmitted through the bites of infected Aedes mosquitoes, although mother-to-child and sexual transmission have also been described. The presence of Zika virus in semen after infection seems to be not uncommon, but the duration of viral persistence has not been well-determined. METHODS: Molecular, serological and cell culture methods were used for the diagnosis and follow up of a case of Zika virus infection imported from Venezuela. Serial samples of serum, urine and semen were analyzed to investigate the persistence of the Zika virus. RESULTS: Zika virus was detected in semen samples up to 93 days after the onset of symptoms. CONCLUSIONS: Our results confirm the persistence of Zika virus in semen samples for long periods after infection.