Paratuberculosis is an economically important disease in dairy cows and requires continuous surveillance. The study aimed to investigate the seroprevalence of bovine paratuberculosis (Johne’s disease) in one of dairy farm in Egypt. A total of 964 dairy cattle were blood sampled and examined with an ELISA method. One-hundred fifty-five (16.1%) samples reacted positively. The results revealed that age was significantly associated with the prevalence of paratuberculosis in dairy cattle, particularly in animals over 6 years of age. Furthermore, the lactation period, milk yield and pregnancy had non-significant effect on appearance of paratuberculosis in cattle.