title: Molecular detection of Brucella species in patients suspicious of Brucellosis from Zanjan, Iran.
abstract: Brucella is an intracellular pathogen capable of infecting animals and humans. The aim of this study was to identify Brucella spp in sera of high risk individuals by a polymerase chain reaction (PCR)-based method. A total of 180 patients suspected to have Brucellosis were examined by serological tests. To establish a PCR protocol for diagnosis of active brucellosis, DNA was extracted from the serum samples by using a commercial kit. PCR amplification was done for detection of Brocella DNA using BCSP31 target gene and IS711 locus. The PCR assay showed that an amplicon of 223 bp was obtained in 73.8% (133/180) of the tested sera using primers (B4/B5) derived from a gene encoding the 31-kDa Brucella abortus antigen. In another PCR, an amplicon of 498 bp was obtained in 63.8% (115/180) of the samples using Brucella abortus-specific primers derived from a locus adjacent to the 3'-end of IS711, and also an amplicon of 731 bp was produced in 4.4% (8/180) of the tested samples using Brucella melitensis-specific primers. When the Wright method was used as a gold standard, the sensitivity and specificity of the PCR technique for genus identification were found to be 96 and 80.7%, respectively. However, the sensitivity value obtained with the species-specific PCR method was 82%, and specificity was similar to that previous reported. This is the first report of a high frequency of Brucella abortus in patients suspicious of Brucellosis from the Zanjan province.
authors.affiliation: Department of Microbiology Faculty of Medical and Basic Sciences Islamic Azad University Zanjan Iran Department of Microbiology, Faculty of Medical and Basic Sciences, Islamic Azad University, Zanjan, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Garshasbi
authors.affiliation: Biotechnology Department, School of Pharmacy Zanjan University of Medical Sciences Zanjan Iran Biotechnology Department, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Ramazani
authors.affiliation: Molecular Biology Research Center Baqiyatallah University of Medical Sciences Tehran Iran Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Rahim
authors.initials: R
authors.lastname: Sorouri
authors.affiliation: Biotechnology Department, School of Pharmacy Zanjan University of Medical Sciences Zanjan Iran Biotechnology Department, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran.
authors.forename: Siamak
authors.initials: S
authors.lastname: Javani
authors.affiliation: Department of Microbiology Faculty of Medical and Basic Sciences Islamic Azad University Zanjan Iran Department of Microbiology, Faculty of Medical and Basic Sciences, Islamic Azad University, Zanjan, Iran.
authors.forename: Soheila
authors.initials: S
authors.lastname: Moradi
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D002003:Brucella abortus / Q000235:genetics / Q000302:isolation & purification*; D017347:Brucella melitensis / Q000235:genetics / Q000302:isolation & purification*; D002006:Brucellosis / Q000175:diagnosis*; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D006801:Humans; D007492:Iran; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D044967:Serum / Q000382:microbiology*; D055815:Young Adult
chemical_list: C527376:31-kDa cell surface protein, Brucella abortus; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial
keywords: Brucella abortus; PCR; human Brucellosis; serologic tests