The present study was aimed to understand the molecular epidemiology of Staphylococcus aureus (54 isolates), isolated from 422 milk samples obtained from 108 subclinical mastitis affected cows (CMT positive >= 1+ in at least one quarter). The molecular epidemiology of Staphylococcus aureus was studied using coagulase (coa) gene polymorphism. 16S-23S ribosomal spascer (RS-PCR) polymorphism and Staphylococcal protein A (Spa) typing. Staphylococcus aureus produced 7 coagulase genotypes and 8 RS genotypes respectively. Coagulase genotype GTIII (730 bp) was the most prevalent (35 strains) followed by GTV (900 bp, 7 strains) and GTIV (800 bp. 4 strains), whereas RS genotypes GTA accounted for the highest number of strains (31 strains), followed by GM (11strains), GTH (4 strains) and GTE (3 strains). Coagulase genotype CTIII (730 bp) showed the highest diversity, as isolates within it produced 5 RS genotypes, the majority of them belonging to the RS genotype GTA (29 out of 31 strains). Forty out of 54 Staphylococcus aureus samples isolated in this study were correctly typed by spa typing, and were assigned to 21 known spa types. and one new novel spa type t18462. The study revealed high diversity within Staphylococcus aureus strains, consisting of 7 coagulase genotypes, 8 RS genotypes and 22 spa types.