Brucellosis is a major infectious zoonotic disease affecting animal and human.In animal, the disease is causing great economic losses such as abortion, still birth as well as infertility in both male and female.A cross sectional study carried out from November 2016 to May 2017 to estimate the seroprevalence of brucellosis in sheep and goatsin Benha.A total of 38 animals (20 sheep, 18 goats)were investigated. 19 serum samples (8 from sheep and 11 from goats) subjected for serological examination. The results showed that,5/8(62.5%) and 9/11(81.8%) from sheep and goats, respectively, were positive by Rose Bengal Plat test(RBPT), Buffered Acidified Plat Antigen test(BABA)and Rivanol test (RIV test).Atotal of 34 lymph nodes from seropositive sheep and goats were examined bacteriologically. 3/20 (15%) and 4/14 (28.57%) were culture positive for Brucellarespectively. None of vaginal swabs revealed culture positive results.All isolates were confirmed genetically as Brucella spp. by PCR and all strains identified asB. melitensis by AMOS- PCR.