تعيين بيوتيپ هاي بروسلاملي تنسيس در ايزوله هاي انساني و گوسفندي

Determination of Brucella melitensis biotypes in human and sheep isolates

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 I. 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.