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

Jeffrey T Foster, Richard T. Okinaka, Rita Svensson, Kathryn Shaw, Barun K. De, Richard A. Robison, William S. Probert, Leo J. Kenefic, William D. Brown, Paul S Keim

Research output: Contribution to journalArticle

46 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)296-301
Number of pages6
JournalJournal of Clinical Microbiology
Volume46
Issue number1
DOIs
StatePublished - Jan 2008

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Brucella
Single Nucleotide Polymorphism
Real-Time Polymerase Chain Reaction
Host Specificity
Essential Genes
Zoonoses
Livestock
Genome
Bacteria
Polymerase Chain Reaction
DNA

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

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

In: Journal of Clinical Microbiology, Vol. 46, No. 1, 01.2008, p. 296-301.

Research output: Contribution to journalArticle

Foster, JT, Okinaka, RT, Svensson, R, Shaw, K, De, BK, Robison, RA, Probert, WS, Kenefic, LJ, Brown, WD & Keim, PS 2008, 'Real-time PCR assays of single-nucleotide polymorphisms defining the major Brucella clades', Journal of Clinical Microbiology, vol. 46, no. 1, pp. 296-301. https://doi.org/10.1128/JCM.01496-07
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 S. / Real-time PCR assays of single-nucleotide polymorphisms defining the major Brucella clades. In: Journal of Clinical Microbiology. 2008 ; Vol. 46, No. 1. pp. 296-301.
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