A cross-sectional survey was conducted on serum samples collected from North and South Gondar zones of North Western Amhara Regional State, Ethiopia to determine the sero-prevalence of foot and mouth diseases in bovine species. The samples were processed with the nonstructural protein (NSP) enzyme linked immunosorbent assay (ELISA) that can able to differentiate foot and mouth disease (FMD) naturally infected vaccinated animals. From the total sera tested, the overall sero-prevalence of foot and mouth disease in cattle at the North and South Gondar zones was 14.9% (86/578). The prevalence rate was high in North Gondar, 17.8% (66/370), as compared to 9.6% (20/208) in South Gondar and the difference was statistically significant (χ² =7.108, p=0.008). Highest sero-prevalence was observed at Metema (62.5%), Quara (46.7%) and Alefa Takusa (34.9%) districts of North Gondar zone, which are bordering with Sudan. The difference among the districts is also statistically significant (χ²=141.115, p=0.0001). There was also a significant association between seropositivity and age groups (χ²=9.483, P=0.009) but there was no significant association between the seropositivity and sex (χ²=0.623, p=0.430). This information on sero surveillance of foot and mouth disease in cattle is important for further epidemiological studies towards developing effective foot and mouth disease control strategies, particularly in these areas where animal movement is not restricted.Keywords: Cattle, Seroprevalence, FMD, North West Amhara Region, 3ABC ELISA