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Objective: A suspected Brucella (B.) strain(GZZA), isolated from a case of anti-Brucella antibody positive patient was identified and its' genetic characteristics was analyzed, to provide etiologic basis for the confirmation of patient in Guizhou province., Methods: Conventional methods and polymerase chain reaction(PCR)were used to identify the bacteria strain, with genetic characteristics analyzed by MLVA-16., Results: The bacteria strain was identified as B. melitensis biovar 3 under the conventional and PCR methods. Results from the MLVA-16 analysis indicated that the bacteria strain was closely clustered with B. melitensis biovar 3, and differences of repeated numbers at VNTR loci bruce42, bruce04, bruce09 and bruce16 were also displayed., Conclusion: Both traditional and molecular methods to identify one bacteria strain isolated from the human patient as B. melitensis biovar 3 and the genetic characteristics of the strain was closely related to that of B. melitensis biovar 3. Differences of repeated numbers at part of VNTR loci were also showed. The results of this study provided etiologic evidences for the confirmation of Brucella infection of the patient, also providing scientific basis for the control and prevention of Brucellosis in Guizhou province.