Molecular Typing of Brucella melitensis and B. abortus From Human Blood Samples Using PCR-RFLP Method

Brucella is an intracellular parasite of the disease brucellosis throughout the world. Although several molecular typing methods are introduced to find DNA polymorphism that is able to identify Brucella species and biovars, but among these methods, detection of polymorphisms by PCR–RFLP has several advantages including the easy implementation, interpretation and the ease of use for large quantities of samples.. In the current study, the technique was used for molecular typing of Brucella abortus and B. melitensis that was isolated from human blood samples.. Blood samples of 160 patients were transferred to Kerman clinical centers with chief complain of acute brucellosis and showed high blood serum level (about 1.80). Their DNAs were extracted by Phenol chloroform method, and the PCR was optimized by using the fragments of designed primers for omp2a and omp2b. Therefore PCR products were restricted by restriction endonuclease as PstI and Hinf1. Finally they were electrophoresed for analyzing the digestion results on agarose gels (2%).. In 160 blood samples that were studied with PCR technique, 52 cases obtained bands of 1100 bp for omp2a locus and 1200 bp for omp2b locus from within 52 positive samples by PCR–RFLP method 25 cases (48%) were positive out of which 56% were B. melitensis biovar1 and 44% were B. abortus biovars of 3, 5, 6 or 9.. The results of this study showed that PCR–RFLP technique was a fast and applicable method especially for separation, detection and differentiation between species of B. melitensis and B. abortus biovars in blood sample. Also the presented data showed that B. melitensis biovar 1 was the prevalence biovar..