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Real-time PCR assays of single-nucleotide polymorphisms defining the major Brucella clades

Foster, Jeffrey T. and Okinaka, Richard T. and Svensson, Rita and Shaw, Kathryn and De, Barun K. and Robison, Richard A. and Probert, William S. and Kenefic, Leo J. and Brown, William D. and Keim, Paul (2008) Real-time PCR assays of single-nucleotide polymorphisms defining the major Brucella clades. Journal of Clinical Microbiology, 46 (1). pp. 296-301. ISSN 1098-660X


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Publisher’s or external URL: http://dx.doi.org/10.1128/JCM.01496-07


Members of the genus Brucella are known worldwide as pathogens of wildlife and livestock and are the most common organisms of zoonotic infection in humans. In general, brucellae exhibit a range of host specificity in animals that has led to the identification of at least seven Brucella species. The genomes of the various Brucella species are highly conserved, which makes the differentiation of species highly challenging. However, we found single-nucleotide polymorphisms (SNPs) in housekeeping and other genes that differentiated the seven main Brucella species or clades and thus enabled us to develop real-time PCR assays based around these SNPs. Screening of a diverse panel of 338 diverse isolates with these assays correctly identified each isolate with its previously determined Brucella clade. Six of the seven clade-specific assays detected DNA concentrations of less than 10 fg, indicating a high level of sensitivity. This SNP-based approach places samples into a phylogenetic framework, allowing reliable comparisons to be made among the lineages of clonal bacteria and providing a solid basis for genotyping. These PCR assays provide a rapid and highly sensitive method of differentiating the major Brucella groups that will be valuable for clinical and forensic applications.

Item Type: Article
Additional Information: Article and Erratum with corrections attached to citation.
ID number or DOI: 10.1128/JCM.01496-07
Related URLs:
Subjects: Q Science > QR Microbiology
NAU Depositing Author Academic Status: Faculty/Staff
Department/Unit: Research Centers > Center for Microbial Genetics and Genomics
Date Deposited: 19 Oct 2015 16:33
URI: http://openknowledge.nau.edu/id/eprint/835

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