Molecular diagnosis of bovine tuberculosis plays an essential role in the epidemiological knowledge of the disease. Bovine tuberculosis caused by represents a risk to human health. This study aimed to perform the genotypic characterization of isolated from bovines diagnosed as tuberculosis from dairy herds in the state of Pernambuco, Brazil. Granulomas from 30 bovines were sent for microbiological culture, and colonies compatible with spp. were obtained in at least one culture from 17/30 granulomas. All isolates were confirmed to be by and 24 MIRU-VNTR typing. While characterized the isolates as SB0121, SB0295, SB0852, SB0120, and an unclassified genotype, 24 MIRU-VNTR rendered two clusters of two isolates each and 13 unique profiles. ETR-A showed higher discriminatory power, and (ETR-B, ETR-C, MIRU16, MIRU27, and QUB26) showed moderate allelic diversity. This is the first study on the genetic variability of the infectious agent cause of bovine TB in Pernambuco and demonstrates variability of strains in the state. Thus, it corroborates the importance of this microorganism as agent of bovine tuberculosis and its zoonotic potential, this epidemiological tool being a determinant in the rigor of the sanitary practices of disease control in dairy herds.