This study aimed to diagnose Lumpy skin desease by isolation and identification of LSD from suspected naturally infected cattle and to study its effect on immuno-biochemical parameters, inflammatory and oxidative stress markers. The study was conducted on 25 cattle, 1-3 years age from two cities in the Sharkia governorate. Ten clinically healthy cattle represented the control group and fifteen suspected diseased cases by LSD represented the infected group. The clinical signs of suspected cases showed anorexia, skin nodules all over the body, fever above 40˚C, edema in forelimbs and dewlap, enlarged superficial lymph nodes, retarded milk production, and abortion in some cases. The virological examination of LSDV from the samples of skin nodules was done. Isolation from skin nodules was done on MDBK cell line and embryonated chiken egg through chorioallantoic membrane route followed by identification using fluorescence technique . Molecular studies including phylogenetic analysis for three positive isolates  showed similarity to previously locally isolated strains in Egypt and close relation to strains of SPPV. Immuno-biochemical results revealed a significant increase in aspartate aminotransferase , alanine aminotransferase , alkaline phosphatase , Malondialdehyde , Tumor necrosis factor , Interleukin4,immunoglobulin M  and immunoglobulin G . Meanwhile, a significant decrease in total protein, albumin, globulin, Glutathione peroxidase, phagocytic assay, calcium, inorganic phosphorous, sodium, and potassium with a non-significant change in magnesium was seen when compared to control ones. It could be concluded that the infection with LSD  in cattle induce severe  effects on immuno-biochemical, antioxidant parameters, and inflammatory markers causing great economic loss