This study was conducted to determine if there is a relationship between vitiligo and ABO blood groups, the Rhesus (Rh) factor, thyroid stimulating hormone (TSH) and vitamin D. For vitiligo analysis, two hundred subjects participated in this study, 100 vitiligo patients and 100 control cases (without vitiligo). ABO blood grouping and Rh typing were tested by a slide method. TSH testing involved 80 vitiligo patients and 80 controls (without vitiligo) and the hormone was analyzed by separating the serum in a centrifuge for two minutes and the results were obtained by Beckman fully automatic analyzer. For vitamin D, 50 vitiligo patients and 50 healthy people (without vitiligo) were included. The data on vitamin D were obtained from private laboratory services. Statistical analysis was performed using IBM SPSS version 26. P<0.05 was considered significant. Most patients with vitiligo had a significantly lower level of serum vitamin D compared with controls (p-value < 0.05), while no statistically significant difference in TSH serum levels between vitiligo cases and controls, was found (p-value > 0.05). Furthermore, despite showing that subjects with blood group O are more susceptible to vitiligo as compared to other groups, there was no significant association of vitiligo with ABO blood groups (p-value > 0.05). Similarly, the incidence of Rh positive and Rh negative was not statistically different between the two groups (p-value > 0.05). This study showed that vitiligo patients are often vitamin D deficient. This study highlights the need to evaluate vitamin D status in vitiligo patients to improve the level of skin pigment loss. It remains unknown whether vitamin D deficiency causes vitiligo. However, a collection of larger sample sizes of different ethnicities should be required to achieve a precise conclusion.