Rapid identification and discrimination of Brucella isolates by use of real-time PCR and high-resolution melt analysis.
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.
Winchell
Jonas M
JM
National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA. jwinchell@cdc.gov
Wolff
Bernard J
BJ
Tiller
Rebekah
R
Bowen
Michael D
MD
Hoffmaster
Alex R
AR
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
D017931:DNA Primers; D004269:DNA, Bacterial