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Six species have classically been recognised within the family <jats:italic>Brucella<\/jats:italic> largely based on a combination of classical microbiology and host specificity, although more recently additional isolations of novel <jats:italic>Brucella<\/jats:italic> have been reported from various marine mammals and voles. Classical identification to species level is based on a biotyping approach that is lengthy, requires extensive and hazardous culturing and can be difficult to interpret. Here we describe a simple and rapid approach to identification of <jats:italic>Brucella<\/jats:italic> isolates to the species level based on real-time PCR analysis of species-specific single nucleotide polymorphisms (SNPs) that were identified following a robust and extensive phylogenetic analysis of the genus.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>Seven pairs of short sequence Minor Groove Binding (MGB) probes were designed corresponding to SNPs shown to possess an allele specific for each of the six classical <jats:italic>Brucella<\/jats:italic> spp and the marine mammal <jats:italic>Brucella<\/jats:italic>. Assays were optimised to identical reaction parameters in order to give a multiple outcome assay that can differentiate all the classical species and <jats:italic>Brucella<\/jats:italic> isolated from marine mammals. The scope of the assay was confirmed by testing of over 300 isolates of <jats:italic>Brucella<\/jats:italic>, all of which typed as predicted when compared to other phenotypic and genotypic approaches. The assay is sensitive being capable of detecting and differentiating down to 15 genome equivalents. We further describe the design and testing of assays based on three additional SNPs located within the 16S rRNA gene that ensure positive discrimination of <jats:italic>Brucella<\/jats:italic> from close phylogenetic relatives on the same platform.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>The multiple-outcome assay described represents a new tool for the rapid, simple and unambiguous characterisation of <jats:italic>Brucella<\/jats:italic> to the species level. Furthermore, being based on a robust phylogenetic framework, the assay provides a platform that can readily be extended in the future to incorporate newly identified <jats:italic>Brucella<\/jats:italic> groups, to further type at the subspecies level, or to include markers for additional useful characteristics.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1471-2180-8-86","type":"journal-article","created":{"date-parts":[[2008,6,3]],"date-time":"2008-06-03T06:14:05Z","timestamp":1212473645000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":64,"title":["Rapid identification of Brucella isolates to the species level by real time PCR based single nucleotide polymorphism (SNP) analysis"],"prefix":"10.1186","volume":"8","author":[{"given":"Krishna K","family":"Gopaul","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark S","family":"Koylass","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine J","family":"Smith","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrian M","family":"Whatmore","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2008,6,2]]},"reference":[{"key":"519_CR1","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1051\/vetres:2005003","volume":"36","author":"J Godfroid","year":"2005","unstructured":"Godfroid J, Cloeckaert A, Liautard JP, Kohler S, Fretin D, Walravens K, Garin-Bastuji B, Letesson JJ: From the discovery of the Malta fever's agent to the discovery of a marine mammal reservoir, brucellosis has continuously been a re-emerging zoonosis. 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