Brucella transmission and epidemiology depend on infecting species and biovar. Therefore, exact identification of the Brucella is important to design correct control and treatment strategies. In this study, we examined presence of otherBrucellae in Isfahan. One hundred twenty Brucella isolates were collected and genomic DNA was extracted from them. OMP2A fragment of all isolates were amplified using a pair of specific primers and the PCR products were lectrophoresed and stained with EtBr. These PCR products were then restricted using PstI restriction endonuclease. The PCR products of all isolates had the same size of 1100bp. The banding pattern of PCR-RFLP for all of the isolates were similar to banding pattern of the Brucella melitensis biotype 1 except for 5 samples that demonstrated banding pattern similar to B. abortus. Based on our results, it is clear that biotype 1 of the B. melitensis is not the only Brucella present in Isfahan and now B. abortus is also present in our area. These results are very important in planning for the control of the disease as well epidemiology and even treatment of the patients.

Background and aim: Brucellosis is one of the most important zoonsis. Because of the importance of this disease and the effect of strain variation on strategic planning for the control of this disease, sheep and human Brucella isolates were evaluated for strain variation. Methods: Five human blood Brucella isolates and 25 sheep embryo Brucella isolates were collected from hospitals and veterinary centers in Isfahan. Total genomic DNA was extracted from the collected samples using method of SDS and proteinase K treatment, phenol/chloroform extraction and ethanol precipitation. OMP2A and omp2b fragments of all isolates were amplified using 2 pairs of specific primers (OMP2A R, F and omp2b R, F) and the PCR products were electrophoresed and stained. These PCR products were then restricted using Alu I, Hinf I and Taq I restriction endonuclease.Results: The PCR products of all human and sheep isolates had the same size 1100bp for OMP2A and 1200bp for omp2b. The banding pattern of PCR-RFLP for all of the isolates was similar to banding pattern of the Brucella melitensis biotype 1.Conclusion: Based on the banding pattern of PCR-RFLP of the OMP2A and omp2b fragments, it can be concluded that all the studied samples in Isfahan are Brucella melitensis biotype 1. This study should be repeated regularly. This will inform us if new species or biotype of Brucella have entered to the region. This is important in planning for the control of the disease.

Brucellosis is a worldwide zoonosis causing reproductive failures in livestock. It is shown that although the vaccine can prevent abortion, it does not provide complete protection against infection. So that vaccination of ewes with Rev.1 biotype can be a source of cattle and even human Brucellosis. The aim of this study was to evaluate possibility of Brucella cross-infection in cattle from Brucella vaccine Rev.1 biotype in Iran. After Brucella vaccination in cattle and ewes in tradition farms in which cattle and sheep are kept close together, 70 aborted cattle fetus samples were collected. The species of Brucella were characterized ant the type of Brucella was studied, by PCR-RFLP method of Pst1 restriction digest of part of OMP2A gene. Of the 70 studied samples, 50 (about 70%) were infected with Brucella Spp. Banding pattern of PCR-RFLP of OMP2A gene demonstrated that in 2 of the samples (4%) the source of infection was Rev.1 vaccine. No sample was infected with Rb51 and S19 vaccine strains, which are specific for cattle vaccination. Our results demonstrated that Rev.1 vaccine biotype can cause Brucella infection in cattle. Therefore in places that cattle and sheep are kept close together, ewe vaccination (Rev.1) can be a source of Brucella abortion in cattle. This study has highlighted some of the potential hazards associated with the use of the Rev.1 vaccine in national control programs.

Background: Brucellosis is a zoonotic disease that causes major economic and public health problems. It is one of the most important diseases in humans and domestic animals. Hence, the exact identification of Brucella spp. is important for strategies of treatment and control. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is one of the molecular techniques characterized by amplification of deoxyribonucleic acid (DNA) sequence and restriction enzyme digestion. Objectives: This study aimed at identifying genetic polymorphisms of OMP2A genes among 90 Brucella isolated from humans and animals, using the PCR-RFLP method. Methods: Ninety Brucella spp. isolated from humans and animals in two different regions of Iran were used in this study. Biochemical tests and the Brucella OMP2A (1100 bp) gene-PCR was used for identification of Brucella isolates. Polymerase Chain Reaction products were digested by restriction endonuclease enzyme pstI and gene sequencing analysis was carried out for molecular typing of Brucella strains. Therefore, genetic relatedness was revealed by a dendrogram. Results: Analysis of the90Brucella strains by biochemical tests, PCR, and PCR-RFLP methods with PstI enzymeandOMP2Asequencing showed four unique RFLP Profiles (P1-P4). Seventy-nine (87. 8%) of the Brucella isolates belonged to B. melitensis strain 20236. From 30 animal isolates, nine (30%) belonged to B. melitensis biovare1 and two (6. 6%) to B. abortus strain. According to the RFLP dendrogram, group 1 and 2 had higher genetic relatedness similarity. Conclusions: The results showed B. melitensis strain 20236 was the predominant strain among human and animal Brucella isolates. Likewise, according to dendrogram results, the PCR-RFLP technique was not able to separatehumanand animal species of B. melitensis from B. abortus.

Brucellosis has always high incidence in Iran, especially we are facing a new outbreak of the disease since 2002. Bakhtiiari tribe sheep enter to the central region of Iran from Iraq border, and therefore can easily introduce new species and/or biotypes of Brucella SPP to these regions. The aim of this study was to determine the species and biotypes of Brucellae that are present in the central region of Iran. We studied 150 Brucella isolates from aborted fetuses of sheep and cow. After identification of species of the bacteria, DNA was extracted and then parts of OMP2A and omp2b genes of the bacteria were PCR amplified. The PCR products were digested by PstI restriction enzyme and the produced patterns of RFLP were analysed. Molecular technique of RFLP results demonstrated that only 135 samples were infected with Brucella melitensis biotype I and 15 samples demonstrated RFLP pattern similar to B. abortus (uncharacterized biotypes) or B. melitensis SPP (biotypes other than biotype 1). Our results demonstrated that although B. melitensis biotype I is the main Brucella in central region of Iran but about 10% of the aborted fetuses were infected with different biotypes of either B. abortus or B. melitensis. We are going to further characterize these biotypes in our next study.

Aim and Background: Brucellosis is one of the most important diseases of zoonosis and is very important in many countries, including Iran, from a health and economic point of view. In this study, the Genotyping of Brucella bacteria isolated from samples of patients suspected of brucellosis was evaluated by PCR RFLP in Tehran. Material and methods: Out of 453 Blood samples suspected of brucellosis of patients who had referred to laboratories in Tehran were inoculated in Castanida culture media and stored at 37 °,C for 21 days. DNA was extracted from colonies that had grown on the surface. Brucella OMP2A gene was amplified by PCR using a specific primer. PCR products were analyzed with Pst1 restriction enzyme, digestion and RFLP patterns. Results: Out of the total samples, it grew in 10 colonies and was confirmed by Brucella by PCR method. In RFLP test, in 8 specimens, Brucella melitensis and in 2 specimens, Brucella abortus were detected. Conclusion: There is brucellosis in our country, infections are also observed in some villages, and among these cases, Brucella melitensis is more common than other species and with the help of molecular methods, in the shortest possible time, high accuracy and sensitivity and with less risk, the presence of different diseases and species of Brucella can be detected.

Objective(s): Rapid and accurate detection of Brucella abortus and Brucella melitensis from clinical samples is so important because antibiotic treatment has major side effects. This study reveals a new method in detection of clinical samples of brucellosis using real‐time PCR and high‐resolution melt (HRM) curve analysis.Materials and Methods: 160 brucellosis suspicious samples with more than 1/80 serum antibody titers were collected and the results were compared with the RFLP method. In order to amplify the sequences for HRM analysis, vdcc, int‐hyp and glk and for RFLP, OMP2A and omp2b with PstI and Hinf1 restriction endonuclease were used. At last, the accuracy and specificity of the two methods were compared with each other.Results: Out of these 160 samples, multiplex real time PCR showed 108 positive samples (67.5%), including 56%B. melitensis and 44% B. abortus; whereas in PCR‐RFLP 52 out of 160 samples were positive, where recognition of two species were accordant with HRM analysis, separation was based on the size of the amplified fragment. Using the designed primers and performing the assay, we confirmed this method to be much faster and have lower cost with more than 99% accuracy compared to methods such as RFLP.Conclusion: The present study showed that this technique, which scans gene segments and creates an analysis pattern for detection of clinical samples, is useful and more dominant compared with PCR‐RFLP.Thus, this method can be used for brucellosis detection, and clinical and epidemiological research.