Key words:Brucella species, RBPT, MRT, bcsp-31 gene Brucellosis is a worldwide zoonotic disease that cause major public health hazard in Egypt. The present work was conducted to investigate the prevalence of bovine brucellosis on the basis of serological test RBPT and MRT in serum and milk samples of lactating dairy animals, also isolation of brucella organism from milk samples, PCR amplification was done for detection of Brucella abortus DNA using bcsp-31 target gene. 219 serum samples from sheep and goat and 119 milk samples from cow, sheep and goat were collected from governmental and private farm. The prevalence of brucellosis by RBPT were 34.5 and 61.4% in serum samples of sheep and goat while by MRT were 2.33, 13.8 and 10% in milk samples of cow, sheep and goat, respectively. The same prevalence was obtained when milk samples were cultured on the Brucella agar medium. A specific PCR assay targeting the bcsp-31 gene within the genome of Brucella species has been used and gave an amplicon of 223 bp in three strain of isolated brucella. It is concluded that RBPT and MRT used as screening tests for detection the prevalence of brucella species in serum and milk samples, PCR assay consider as a useful tool for detection Brucella species in milk.

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