Abstract:

In this study, we designed and used 2 different type-specific polymerase chain reaction (PCR) methods for the detection of Brucella abortus. Materials and methods: There were 2 different primer sets (B4/B5 and AF/AR) selected and used for the amplification of 2 different genes that are present in all biovars of Brucella species, including bcsp31, encoding 31-kDa immunogenic cell surface proteins, and omp25, which is known to be one of the virulence factors encoding outer membrane proteins of 26–23 kDa. Results: The results showed that 273 (81.7%) and 317 (94.9%) out of 334 milk samples were positive for brucellosis, as detected by either both or one of the primer sets used in the present study, respectively. The detection limit of PCR assays for Brucella in milk samples was determined as 5 pg DNA for both of the primer sets. Conclusion: These results suggest that the use of the specific PCR assay with the primer sets used in the study is a rapid, reliable, and accurate technique in comparison to traditional and conventional methods for the detection and diagnosis of Brucella spp. in milk samples. Kew words: Brucella abortus, bovine milk, bcsp31-PCR, omp25-PCR

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