One-step RT-PCR for detection of Zika virus

Background: Zika virus (ZIKV) is an emerging mosquito-borne flavivirus circulating in Asia and Africa. Human infection induces an influenza-like syndrome that is associated with retro-orbital pain, oedema, lymphadenopathy, or diarrhea. Diagnosis of Zika fever requires virus isolation and serology, which are time consuming or cross-reactive. Objective: To develop a one-step RT-PCR assay to detect ZIKV in human serum. Study design: An assay targeting the envelope protein coding region was designed and evaluated for its specificity, detection limit, repeatability, and capacity to detect ZIKV isolates collected over a 40-year period from various African countries and hosts. Results: The assay's detection limit and repeatability were respectively 7.7 pfu/reaction and 100% in serum and L-15 medium; none of 19 other flaviviruses tested were detected. Conclusions: The assay is rapid, sensitive, and specific to detect ZIKV in cell culture or serum, but needs to be validated for diagnosis using clinical samples