Abstract: A pair of primers were designed according to the sequence of specific Brucella gene of Outer membrane protein(OMP31),the OMP31 gene was amplified by PCR and cloned into pMD18-T vector, and then the gene sequence was detected. The results showed that the target gene band at a length 602 bp was amplified by PCR, with a homology of 99.7% to the gene sequence reported in GenBank. The experiments had proved that PCR assay possessed a high specificity, DNA bands couldn't amplified in other bacterial except Brucella. And the sensitivity test results indicated that PCR assay was more sensitive, which could detect OMP31 with only 0.9 pg/μL DNA. The successfully construction of the PCR assay provides strongly technical support for detection, identification and epidemiological investigations of bovine brucellosis.

Key words: bovine; Brucella; PCR; detection