Bovine papular stomatitis (BPS) is a widespread epitheliotropic viral disease affects ruminants of all ages and considered a serious risk to public health. In Assiut governorate, there is little information on BPS despite their financial losses. Thus, the goal of this study was to confirm diagnosis on a molecular base, describe the clinical findings of the disease, and study the relationship between some factors that may play a role in the spread of the disease. The present investigation was conducted on 39 cattle and 11 buffaloes that belonged to different villages in Assiut governorate, Egypt. Whole blood and oral lesions swabs samples were taken for laboratory testing. Semi-nested polymerase chain reaction (semi-nested PCR) had been used for detection of BPS virus (BPSV). The viral DNA was detected in 32 cattle and 8 buffaloes. The result of clinical examination indicated that the clinical signs of BPS were oral (ulcers in gum, palate, papillae & tongue), commissure, muzzle, and nostrils lesions with one or more of other clinical signs such as fever, diarrhea, enlarged superficial lymph nodes, corneal opacity, respiratory distress and skin lesions in studied animals. Studying the effect of some factors on the spread of the disease revealed that the prevalence was significantly higher in middle areas of the governorate than in other areas. To lower the prevalence of BPSV, it is recommended to avoid eating hard hay and reeds that cause small abrasion in oral cavity and aid increasing frequency of BPSV infection.