الفهرس 
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Abstract
ABSTRACT
Brucellosis is one of the endemic diseases in Egypt, because of these when problems associated with abortion is present, the main cause suspected is brucellosis, so its serological diagnosis is applied. This study aimed to estimate the serological prevalence of the disease in 7 Different Governorates in Egypt (Giza, Fayoum, Benisuef, El-Menia ,Assiut ,Sohag and Qena.). , isolation and bio typing of Brucella species confirmed by PCR. Study the validity of PCR and 16SrRNASequence for diagnosis of Brucella infection. A total of 1857 samples were collected including 1531 serum, 148 milk, 58 lymph nodes,58 spleen samples, 58 liver samples and 4 aborted foeti from cattle in 7 Governorates in Egypt. Serological tests; Rose Bengal Plate Test (RBPT), Buffered acidified plate antigen (BAPA) test (as screen tests), modified standard tube agglutination (MSTA) and indirect ELISA were applied on positive serum samples for (RBPT). Brucella was isolated and identified from milk, lymph nodes, Spleen and aborted foeti. The results detected 19 Brucella isolates from (aborted foeti 1, milk 8, lymph nodes 8 and spleen 2) were detected and identified as Br..melitensis biovar3. The results of RBPT, BAPA, MSTA and indirect ELISA tests were 21.8%, 23.7%, 80.2%, and 89.8% respectively. MSTA and indirect ELISA applied on positive sera of RBPT. Multiplex PCR was applied as a confirmation and rapid detection of B. melitensis isolates. all isolates showed positive results with oligonucleotide primer that amplified a 731bp fragment confirmed as B. melitensis. one of the isolates was sequenced and identified as Br. melitensis bv.3 strain by applied The 16SrRNA gene (Gene Bank accession No.MT 378400). On sequencing, the Nucleotide sequence alignment of obtained sequences with other Brucella strain indicated that the obtained isolate have high identity with Br. melitensis biovar 3.
Key words: Brucella, Isolation, Identification, Serology, PCR.