A sheep flock contains 600 head of Al-Barky ewes suffered from: Icterus, bloody urine, death two days post symptoms appearance and abortion in 6 pregnant ewes. Thesymptoms appeared on 46 case and 6 ewes aborted and mortality rate was44 case dead. The results of examined samples showed that the isolation of leptospira from 5 out of 52 (9.62 %) animals samples (blood, urine and necropsy kidney tissue samples), the isolates were 2 from blood samples (6.7 %), 2 from kidney samples (6.7 %) and 1 from urine samples (10 %). The obtained leptospira isolates from infected ewes were examined by PCR for identification of leptospires, using universal primers set (lig1 / lig2,) that amplify the genomic DNA of all pathogenic Leptospiraserovar only, the most predominant seropositivity using MAT was detected against L.int.icterohaemorrhagiae (57.69%), followed by L.int.grippotyphosa (28.85%) and L.int.pomona (13.46%). The highest titer of seropositivity was detected against serovarL.int.icterohaemorrhagiae (1:1600) while L.int.grippotyphosa and L.int.pomona reached up to (1:400).