title: Simultaneous detection and differentiates of Brucella abortus and Brucella melitensis by combinatorial PCR.
abstract: OBJECTIVE
  To evaluate simultaneous detection and differentiates of Brucella abortus (B. abortus) and Brucella melitensis (B. melitensis) through the combinatorial PCR method.
  
  
  METHODS
  This study was designed using three primers that could simultaneously identify and differentiate two major species of pathogenic Brucella in humans and animals. Identification and differentiation of each species using the size of the PCR product were determined. To determine the specificity of the method, bacteria close to the genus Brucella were used. Finally, to confirm PCR products, In addition to the products sequence, RFLP was performed on PCR products using restriction enzymes.
  
  
  RESULTS
  The method of optimized combinatorial PCR in this study could simultaneously detect and differentiate B. abortus and B. melitensis with high specificity and sensitivity in clinical samples. Differentiation of species is based on the resulting bands; therefore, the band 494 bp for B. abortus and 733 bp for B. melitensis were obtained. RFLP and sequencing results confirmed PCR results.
  
  
  CONCLUSIONS
  The results of this study shows that without routine diagnostic methods such as culture and serology tests, using the molecular method of combinatorial PCR, important species of Brucella can be simultaneously identified and differentiated in clinical samples.
authors.affiliation: Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. rmirnejadreza@yahoo.com
authors.forename: Reza
authors.initials: R
authors.lastname: Mirnejad
authors.forename: Reza Hosseini
authors.initials: RH
authors.lastname: Doust
authors.forename: Reza
authors.initials: R
authors.lastname: Kachuei
authors.forename: Seied Mojtaba
authors.initials: SM
authors.lastname: Mortazavi
authors.forename: Mehdi
authors.initials: M
authors.lastname: Khoobdel
authors.forename: Ali
authors.initials: A
authors.lastname: Ahamadi
mesh_terms: D000818:Animals; D001483:Base Sequence; D002003:Brucella abortus / Q000235:genetics / Q000302:isolation & purification*; D017347:Brucella melitensis / Q000235:genetics / Q000302:isolation & purification*; D002006:Brucellosis / Q000175:diagnosis* / Q000382:microbiology*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial