Theileria orientalis (T. orientalis) is a benign species of Theileria that is found all throughout the world, including Egypt. The purpose of this work was to determine the current status of T. orientalis infection in cattle, as well as to define T. orientalis genetic diversity using the major piroplasm surface protein (MPSP) genes. In 2020, blood samples from 500 asymptomatic cattle were collected, which live in four different governorates at northern Egypt and examined using PCR assay based on MPSP gene. Overall, 44 blood samples (8.8%, 44/500) were positive for T. orientalis, and the highest prevalence rate was observed in Kafer ElSheikh while it was relatively low in Gharbia governorate. Moreover, based on multivariable analysis, the risk of infection with T. orientalis infection was higher in mixed breed cattle and in animals of age group more than 3 years compared to other. Additionally, absence of acaricide application, infestation with ticks, and communal grazing system were identified as potential risk factors for T. orientalis infection. Phylogenetic analysis revealed that the isolates studied belonged to two MPSP genotypes, types 1 and 2. These findings reveal that the MPSP genotypes discovered in the present work displayed genetic variation that was linked to geographic area. Therefore, further research should be focused on developing an effective T. orientalis monitoring and preventive program.