Lumpy skin disease virus (LSDV) was isolated from skin biopsies collected from clinically infected buffaloes at Kaluobia governorate. The virus was isolated on chorio-allantoic membrane (CAM) of embryonated chicken eggs (ECEs) and identified by agar gel precipitation test (AGPT) and indirect fluorescent antibody technique (IFAT) using specific hyper immune serum against LSDV. Further investigations were carried out using electron microscopic (EM) examination and polymerase chain reaction (PCR) for attachment protein gene confirmed LSDV isolate with specific amplified product 192bp