title: Rapid identification and discrimination of Brucella isolates by use of real-time PCR and high-resolution melt analysis.
abstract: Definitive identification of Brucella species remains a challenge due to the high degree of genetic homology shared within the genus. We report the development of a molecular technique which utilizes real-time PCR followed by high-resolution melt (HRM) curve analysis to reliably type members of this genus. Using a panel of seven primer sets, we tested 153 Brucella spp. isolates with >99% accuracy compared to traditional techniques. This assay provides a useful diagnostic tool that can rapidly type Brucella isolates and has the potential to detect novel species. This approach may also prove helpful for clinical, epidemiological and veterinary investigations.
authors.affiliation: National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA. jwinchell@cdc.gov
authors.forename: Jonas M
authors.initials: JM
authors.lastname: Winchell
authors.forename: Bernard J
authors.initials: BJ
authors.lastname: Wolff
authors.forename: Rebekah
authors.initials: R
authors.lastname: Tiller
authors.forename: Michael D
authors.initials: MD
authors.lastname: Bowen
authors.forename: Alex R
authors.initials: AR
authors.lastname: Hoffmaster
mesh_terms: D000818:Animals; D015373:Bacterial Typing Techniques; D002002:Brucella / Q000145:classification* / Q000302:isolation & purification*; D002006:Brucellosis / Q000175:diagnosis* / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D044366:Transition Temperature
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial