Differentiation of Brucella abortus and B. melitensis biovars using PCR- RFLP and REP-PCR

Background: Brucellosis is one of the common zoonotic diseases of human and livestock. Epidemiologic studies and biovar differentiation provides valuable data to make plans for control and prevention of disease. This study was aimed to differentiate and typing of Brucella spp. and determine the pattern of biovars using REP-PCR and PCR-RFLP methods. Methods: A total of 100 blood specimens suspected to brucellosis were collected from slaughterhouses and different treatment centers in Tehran, Gilan and Hamedan provinces. The conventional culture methods and Multiplex-PCR were used for the detection of Brucella genus and species and also REP-PCR was used to differentiation of Brucella spp. and polymorphisms sequence analysis. To identify the biovar patterns of REP-PCR, PCR-RFLP was used. Results: 83 samples by using Multiplex-PCR were identified as Brucella spp. To analyze the results of REP-PCR, from 83 approved samples of Brucella spp., 72 were Brucella melitensis and 11 were Brucella abortus. Also, in analyzing the results of PCR-RFLP, from 72 B. melitensis, 69 were B. melitensis biovar 1 and three species were from other biovars. Also the obtained patterns were same for 11 B. aborts samples and were from the biovars of 3, 5, 6 and 9. Conclusion: This study was found a significant relationship between polymorphism sequences with B. abortus and B. melitensis biovars. This study also optimized a test for the detection of Brucella biovar with the REP-PCR method that by which, in the shortest possible time, the Brucella spp. and biovars will be separated

Differentiation of Brucella abortus and B. melitensis biovars using PCR- RFLP and REP-PCR Keywords:

Brucella abortus , B. melitensis , polymorphisms sequence , REP-PCR , PCR-RFLP , OMP