The objective of this study was to identify the presence and distribution of virulence genes via multiplex Polymerase Chain Reaction by identification of Streptococcus uberis, one of the agents which cause mastitis in dairy cows. S. uberis strains were identified from samples and stored at -20 °C deep freeze for molecular studies. DNA extraction of the isolated strains was performed according to the procedure with the genomic DNA extraction kit (Fermentas®). The presence of 16S rRNA and virulence-associated genes was examined in all strains identified as S. uberis phenotypically. S. uberis isolation was conducted in 35 (17.5%) of the 200 milk samples. After examination of the identified S. uberis strains by polymerase chain reaction, 16S rRNA gene was detected in a total of 35 isolates (100%) at 1400 bp fragment base pairs. When the virulence genes were examined individually, 32 (91%) strains expressed hasB, 32 (91%) strains expressed gapC, 32 (91%) strains expressed skc, 30 (86%) strains expressed cfu, 30 (86%) strains expressed hasC, 30 (86%) strains expressed hasA 29 (83%) strains expressed sua, 28 (80%) strains expressed oppF, 25 (71%) pauA, 9 (26%) strains expressed Ibp genes. The pauB gene was not identified. When the distribution of virulence genes of S. uberis strains isolated from mastitis cases are examined, it has been shown that the genes which exert capsular formation in terms of the pathogenity of the agent play important role and also the genes involved in the synthesis of serine protease enzymes are distributed in significant proportion.