Human milk is the best nutrient for infants. The donor human milk is stored in a milk bank before pasteurization. However, the human milk is not sterile and could be colonized with different types of bacteria. Many studies have shown S. aureus to be the most prevalent potential pathogen detected in human milk. This study characterized 22 methicillin-resistant and methicillin-sensitive Staphylococcus aureus isolates from raw human milk for the presence of virulence genes and agr type. Moreover, the genotypic as identified characterization was realized. The presence of virulence genes sei, seg, sec, seh, and etb was identified in resistant and sensitive strains. We observed the predominance of agr type II. The presence of SCCmec IV (67%, 4/6) and V (33%, 2/6) characterized resistant strains as CA-MRSA. Endemic lineages detected (ST1635/CC5-t002, ST5/CC5-t002, ST72/CC5-t126, ST1/CC1-t127, ST45/CC45-t065, and ST398/t1451) could be related to epidemic clones, such as USA800/ST5, USA700/ST72, USA400/ST1, USA600/ST45, and ST398. This study made it possible to understand the characteristics of virulence and clonality of some strains that circulate in breast milk in our region. The discovery of human milk colonization by MSSA and MRSA strains with molecular characteristics similar to infectious clones spread globally demonstrates the importance of monitoring strains that can spread and cause serious infections.