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Background and Aim: There are numerous reports of subclinical mastitis cases in Blitar, which is consistent with the region’s high milk production and dairy cattle population. Staphylococcus aureus, which is often the cause of mastitis cases, is widely known because of its multidrug-resistant properties and resistance to β-lactam antibiotic class, especially the methicillin-resistant S. aureus (MRSA) strains. This study aimed to molecular detection and sequence analysis of the mecA gene in milk and farmer’s hand swabs to show that dairy cattle are reservoirs of MRSA strains. Materials and Methods: A total of 113 milk samples and 39 farmers’ hand swab samples were collected from a dairy farm for the isolation of S. aureus using Mannitol salt agar. The recovered isolates were further characterized using standard microbiological techniques. Isolates confirmed as S. aureus were tested for sensitivity to antibiotics. Oxacillin Resistance Screening Agar Base testing was used to confirm the presence of MRSA, whereas the mecA gene was detected by polymerase chain reaction and sequencing. Results: : A total of 101 samples were confirmed to be S. aureus. There were 2 S. aureus isolates that were multidrug-resistant and 14 S. aureus isolates that were MRSA. The mecA gene was detected in 4/14 (28.6%) phenotypically identified MRSA isolates. Kinship analysis showed identical results between mecA from milk and farmers’ hand swabs. No visible nucleotide variation was observed in the two mecA sequences of isolates from Blitar, East Java. Conclusion: The spread of MRSA is a serious problem because the risk of zoonotic transmission can occur not only to people who are close to livestock in the workplace, such as dairy farm workers but also to the wider community through the food chain.