BACKGROUND: Zika virus (ZIKV) is a teratogenic flavivirus that can cause microcephaly. Its main vector, Aedes aegypti, has been previously identified in Saudi Arabia, but no ZIKV infection has yet been reported. Nevertheless, the country is at risk from ZIKV because it receives many travelers throughout the year, including pilgrims from ZIKV-endemic countries. OBJECTIVES: Screen asymptomatic pregnant mothers and their newborns attending a major hospital in the Najran region for subclinical or past infections with ZIKV, using ELISA and RT-PCR. DESIGN: Cross-sectional. SETTING: Najran Maternity and Children Hospital (NMCH). SUBJECTS AND METHODS: All pregnant women admitted to NMCH in labor between November 2016 and July 2017 were included in the study. Clinical and demographic data were collected by pre-validated physician-administered questionnaires. Paired umbilical and maternal serum samples were collected and frozen at -60°C, using ELISA to measure anti-ZIKA IgG and IgM antibodies and RT-PCR to further investigate positive samples. MAIN OUTCOME MEASURES: Maternal and newborn serum antiZIKV IgM and IgG and ZIKV RT-PCR. SAMPLE SIZE: 410 mother-newborn pairs. RESULTS: The median gestational age was 38.5 weeks (range 33-42). Most (n=342, 83.41%) of the women were from Najran city. All of the newborns had normal growth parameters with no congenital malformations. None of the mothers had symptoms suggestive of ZIKV infection; 3 (0.7%) exhibited a low-grade fever (38°C), but did not test positive for anti-ZIKV antibodies. Thirty-five (8.53%) of mothers had travelled inside Saudi Arabia, but none outside the country. Twentyfour (5.85%) mothers tested positive for anti-ZIKV IgM and 52 (12.68%) tested positive for anti-ZIKV IgG, but all infant samples were negative. All seropositive ZIKV IgM were also ZIKV IgG positive, but RT-PCR testing of all seropositive samples was negative. CONCLUSION: Although previous (resolved) ZIKV infection and crossreactivity of the ELISA method with other flaviviruses cannot be excluded, the study found no confirmed cases of acute ZIKV infection. However, given the presence of the vector in Saudi Arabia, the presence of presumptive positive serology and the ongoing risk of ZIKV entry via a regular influx of travelers from endemic areas, we propose that continuous surveillance be conducted for ZIKV as well for other flaviviruses. Larger-scale nationwide studies are strongly recommended to gain a broader view of the potential threat from ZIKV in the country. LIMITATIONS: Small sample size, unavailability of plaque reduction neutralization tests to confirm serology results, and RT-PCR was only conducted on ELISA-positive serum samples, due to resource constraints