In Egypt, outbreak of Lumpy Skin Disease (LSD) was suspected among cattle at Ismailia governorate, May 2016. This work aimed for molecular characterization of Lumpy Skin Disease Virus (LSDV) in skin nodule samples. Suspected virus isolate on embryonated chicken eggs and MDBK cell line was detected successfully by polymerase chain reaction (PCR) using primers specific for the envelope protein gene P32. Nucleotide and amino acid sequence analysis of the envelop protein gene P32 of the isolated KH/LSD/Egypt/2016 revealed complete identity with LSDV-998/Egy/2015; LSDV-956/Egy/2015; LSDV-NI-2490-Neeth and LSDV/Slemani/Kurd/014 with 100% identity. Phylogeny revealed that KH/LSDV/Egypt/2016 present in the same group with LSDV-956/Egy/2015 and LSDV-998/Egy/2015 and is closely related to LSDV-SL1 and LSDV-TK1. It is concluded that there are great homology between the present obtained KH/LSD/Egypt/2016 and other isolates recorded in Egyptian governorates through the last few years, indicated by the high nucleotide sequence identity and close branch distances in the phylogenetic tree