title: Evaluation of genus-specific and species-specific real-time PCR assays for the identification of Brucella spp.
abstract: BACKGROUND
  The identification of Brucella isolates using conventional microbiological techniques is time-consuming and hazardous. We therefore assessed the performance of real-time PCR assays for the identification of members of the genus Brucella to the genus and species level.
  
  
  METHODS
  We evaluated an in-house developed assay and various previously published real-time PCR assays targeting bcsp31, per, IS711, alkB/IS711 and BMEI1162/IS711 using 248 Brucella strains representing the biotypes of all species and a large panel of clinically relevant, phylogenetically related and serologically cross-reacting bacteria.
  
  
  RESULTS
  No misidentification was observed. However, several biotypes of Brucella abortus and Brucella suis were not detected with some of the published assays. The limit of detection varied widely among the assays (16-1600 fg) demonstrating that some assays should not be applied to clinical samples but may help to identify colony material.
  
  
  CONCLUSIONS
  In summary, most of the assays revealed low detection limits and proved to be highly selective for the detection of the genus Brucella and the species that are most relevant for humans. Assays targeting the bcsp31 gene can be recommended to screen for Brucella. Species-specific assays should be consecutively applied confirming the primary diagnosis by a second gene target.
authors.affiliation: Department of Bacteriology, Bundeswehr Institute of Microbiology, Munich, Germany. Sascha.Al-Dahouk@gmx.de
authors.forename: Sascha
authors.initials: S
authors.lastname: Al Dahouk
authors.forename: Karsten
authors.initials: K
authors.lastname: Nöckler
authors.forename: Holger C
authors.initials: HC
authors.lastname: Scholz
authors.forename: Martin
authors.initials: M
authors.lastname: Pfeffer
authors.forename: Heinrich
authors.initials: H
authors.lastname: Neubauer
authors.forename: Herbert
authors.initials: H
authors.lastname: Tomaso
mesh_terms: D001483:Base Sequence; D002002:Brucella / Q000145:classification* / Q000235:genetics; D017931:DNA Primers; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers