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.
Subject(s)
Adolescent , Adult , Aged , Animals , Female , Humans , Male , Middle Aged , Young Adult , Brucella abortus/isolation & purification , Brucella melitensis/isolation & purification , Brucellosis/diagnosis , Molecular Diagnostic Techniques/methods , Polymerase Chain Reaction/methods , Serum/microbiology , Antigens, Bacterial/genetics , Bacterial Proteins/genetics , Brucella abortus/genetics , Brucella melitensis/genetics , DNA Transposable Elements , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Iran , Sensitivity and SpecificityABSTRACT
Aim: The aims of this study were to genotype CYP2C8 in an Iranian population and compare their allelic frequencies with other ethnic groups. Study Site and Duration: Biotechnology Department, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran from June 2012 through May 2013. Methodology: CYP2C8 (*1/*2/*3) allelic variants were determined in 200 unrelated healthy Iranian volunteers by real time-polymerase chain reaction (PCR). Results: A total 156 subjects (78%) were homozygous for CYP2C8*1, six subjects (3%) were homozygous for CYP2C8*2 and 38 subjects (19%) were heterozygous CYP2C8*1/*2. CYP2C8*3 was not detected. Discussion and Conclusion: Genotyping indicated no significant (P>0.05) difference between CYP2C8 allelic variant frequencies in an Iranian compared with Burkina Faso population. The Iranian population’s CYP2C8 allelic variation was significantly (P<0.05) different when compared with populations in Portugal, African-American race to Malaysia, Ghana, Zanzibar, Spain, Czech Republic and Sweden.