Vaginitis is still considered to be a global health problem for females. It is mainly occurred by bacterial vaginosis, trichomonal, candidal, and gonococcal vaginal infections. This study is aimed identify Staphylococcus haemolyticus in females infected with vaginitis and determine some of their virulence genes. A total of 250 vaginal swabs were collected from females with vaginitis. Of the 250 vaginal swabs, 40 isolates were showed bacterial growth of S. haemolyticus. In susceptibility testing, S. haemolyticus isolates were sensitive to some antibiotics, whereas other isolates were resistant. The amplified 16S rRNA gene was in all isolates, using conventional PCR, icaA, blaz, mecA, and SodA genes were detected in 30, 23, 32, and 24 samples, respectively. The sequencing analysis of all local isolates appeared slight mismatching in their sequencing. The PCR products of virulence factors of S. haemolyticus showed slight variations. This may explain different clinical manifestations and the infection severity.