Orf virus infecting sheep and goats belongs to genus Parapoxvirus, subfamily Chordopoxvirinae, family Poxviridae. It causes contagious pustular dermatitis in sheep and goats worldwide, including Egypt. This work aimed for molecular and serological diagnosis of Orf virus in infected sheep and goats during summer season 2020. Ten samples of skin lesions from suspected sheep and goats from different areas of Minufiya governorate including Queisna, Tala, and Ashmoon were subjected to direct detection using PCR primers targeting ORF 045 gene which encodes the late transcription factor (VLTF-1). Positive samples on PCR detection were isolated on Specific Pathogen Free- Embryonated Chicken Eggs (SPF-ECEs) then viral antigen was detected using indirect-fluorescent antibody technique (IFAT) and molecular detection of viral DNA using PCR. Positive results were shown in 7 samples (5/7 of sheep samples and 2/3 of goat samples) using PCR. Virus isolation from positive samples on SPF-ECE that gave positive results by the 3rd passage in 5 of 7 samples (3 sheep isolates and two goat isolates). Signs of inoculated viral samples on CAM of SPF-ECEs showed thickening, edema, and hemorrhage of the membrane. These isolates showed positive results upon identification using IFAT and PCR. It was concluded that Orf virus was circulating among sheep and goats in Minufiya governorate Egypt, during summer 2020.