NDLI: Real-Time PCR Assays of Single-Nucleotide Polymorphisms Defining the Major Brucella Clades▿

Content Provider	PubMed Central
Author	Foster, Jeffrey T. Okinaka, Richard T. Svensson, Rita Shaw, Kathryn De, Barun K. Robison, Richard A. Probert, William S. Kenefic, Leo J. Brown, William D. Keim, Paul
Abstract	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.
Related Links	http://dx.doi.org/10.1128/jcm.01496-07
Ending Page	301
Page Count	6
Starting Page	296
File Format	PDF
ISSN	00951137
e-ISSN	1098660X
Journal	Journal of Clinical Microbiology
Issue Number	1
Volume Number	46
Language	English
Publisher	American Society for Microbiology (ASM)
Publisher Date	2008-01-01
Access Restriction	Open
Rights Holder	American Society for Microbiology (ASM)
Subject Keyword	Microbiology (medical) Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Microbiology (medical)

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