Objective of the study is to identify species appurtenance of Brucella spp. strains, using a complex of restriction and amplification techniques. Materials and methods. Carried out has been revision and clarification of species appurtenance of 17 reference and 48 natural strains, stored in the biobank of the State collection at the RusRAPI “Microbe”, using amplification (“Gen Brucella – identification – RGF“, AMOS-DEL, Bruce-Ladder, Suis-Ladder) and restriction analyses of omp25, omp2a, omp2b loci, applying AluI, EcoRI, Eco32I (EcoRV) ferments. Results and conclusions. Performed molecular identification has confirmed the stated in the profile taxonomic status of 50 Brucella strains, while for 12 isolates species appurtenance has been rectified. New profiles of amplification and restriction have been established for three cultures, which outlines the necessity to continue further investigations. The results obtained fully substantiate the prospects of the complex approach, which implies several amplification systems and preparations, as well as restriction analysis for determination of species and biovar of brucellosis agent strains.