BACKGROUND AND AIM: Rift Valley fever virus (RVFV) is a vector-borne virus that causes RVF in humans and ruminants. The clinical symptoms in humans and animals are non-specific and often misdiagnosed, but abortions in ruminants and high mortality in young animals are characteristic. Since the initial outbreak in the Rift Valley area in Kenya, the disease has spread to most African countries and the Middle East. The presence and epidemiological status of RVFV in humans and animals in Palestine are unknown. This study aimed to investigate the presence and risk factors for RVF seroprevalence in veterinarians, as occupational hazard professionals, and sheep, as highly susceptible animals, in Northern Palestine. MATERIALS AND METHODS: A cross-sectional study was conducted. Data and blood samples of 280 Assaf sheep and 100 veterinarians in close occupational contact with sheep were collected between August and September 2020 using an indirect enzyme-linked immunosorbent assay. RESULTS: No evidence of RVF antibodies was found in any human or animal sample. CONCLUSION: Our results suggest that RVFV has not circulated in livestock in Northern Palestine, yet. Surveillance and response capabilities and cooperation with the nearby endemic regions are recommended. The distribution of competent vectors in Palestine, associated with global climate change and the role of wild animals, might be a possible route for RVF spreading to Palestine from neighboring countries.