Brucellosis is one of the most common zoonotic diseases of animal and human beings. This study aimed to differentiate the Brucella spp. and determines the patterns of biovars by using repetitive element palindromic (REP)-PCR and PCR restriction fragment length polymorphism (RFLP) methods. A total of 100 blood specimens suspected of harbouring brucellosis were collected. Conventional culture methods and multiplex PCR were used for the detection of Brucella genus and species; and REP-PCR was used for Brucella spp. differentiation and polymorphisms sequence analysis. In addition, to identify the biovar patterns of REP-PCR, PCR-RFLP was used. Eighty-three samples were identified as harbouring Brucella spp. by the implementation of multiplex PCR, 72 of which were detected as Brucella melitensis and 11 as B. abortus. Also, through analysing the results of PCR-RFLP, it was found that of 72 B. melitensis samples, 69 were B. melitensis biovar 1 and three species were from other biovars. In addition, the obtained patterns for all of the B. abortus samples were from biovars 3, 5, 6 and 9. This study also optimized a test for the detection of Brucella biovar with the REP-PCR method such that Brucella spp. and biovars could be separated in the shortest possible time.
Brucellosis is one of the most prevalent zoonotic diseases among animals and humans. It is a well known fact that the differentiation and rapid typing of Brucella spp. is crucial for the early detection of infection, prevention of infection progress, and/or introducing treatment solutions. Analyzing the sequences could be an effective method in achieving these purposes. The aim of this study was to analyze palindromic sequences for Brucella spp., differentiation using the rep-PCR method. The authors collected 80 animal samples, which were suspected to brucellosis infection. After the cultivation of Brucella, identification was performed through standard biochemical, microbiological, and IS711 PCR assays. By designing the specific primers for polymorphism sequence, the rep-PCR was performed. The resultant pattern was compared with the obtained patterns of the standard Brucella melitensis and Brucella abortus samples, which showed dissimilar patterns. For this reason, the PCR products were sequenced, and consequently two new patterns were introduced. This rapid and repeatability assay has the ability to potentially differentiate the B. abortus and B. melitensis species, which could be useful in early diagnosis and treatment of patients with brucellosis.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.