RESUMO
We demonstrate that the "HOOF-Print" assay provides high power to discriminate among Brucella isolates collected on a small spatial scale (within Portugal). Additionally, we illustrate how haplotype identification using non-random association among markers allows resolution of B. melitensis biovars (1 and 3). We recommend that future studies use haplotype identification when analyzing multilocus population genetic data to help discriminate among microbial isolates such as Brucella.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Brucella melitensis/genética , Brucella/classificação , Brucella/genética , Genes Bacterianos , Haplótipos , Brucella/isolamento & purificação , Brucelose/microbiologia , Variação Genética , Humanos , Repetições Minissatélites , Modelos Genéticos , Filogenia , PortugalRESUMO
Swine brucellosis is caused by the biovars 1, 2 and 3 of Brucella suis the identification of which up to now relies on microbiological tests lacking adequate specificity together with time consuming and expensive molecular procedures. Based on sequence variation of the omp2b gene, we have developed a four primer set multiplex PCR assay that was tested for polymorphism analysis of B. suis biovars causing brucellosis in swine. The assay exploits the single nucleotide polymorphisms found in omp2b gene of B. suis reference biovars which are conserved in 43 B. suis field isolates from different geographic origins and hosts. Three specific amplification patterns (S1, S2 and S3) were obtained for reference strains of B. suis biovars 1, 2 and 3, respectively. However, some B. suis field isolates identified as biovars 2 or 3 according AMOS-PCR, PCR-RFLP of omp31 and omp2 genes and classical bacteriological methods, resulted also in S1 patterns, limiting the typing usefulness of the method.