Aim: To evaluate the diagnostic and prognostic value of serum 25-hydroxyvitamin D (25(OH) D), ionized calcium and phosphorus in comparison with serum CA15.3 as the most commonly used breast cancer marker. Participants and methods: This study was conducted on 45 breast cancer female patients with recently detected breast cancer before surgery and 45 apparently healthy female controls of matched age, menstrual and socioeconomic status as breast cancer patients group. Serum 25(OH) D, ionized calcium, phosphorus and CA15.3 were measured using readyfor-use commercially available kits. Results: Serum levels of 25(OH) D and ionized calcium in the breast cancer patients group were significantly lower than those of the control group, while serum levels of phosphorus and CA15.3 in breast cancer patients group were significantly higher than those of the control group. The area under the ROC curve for serum ionized calcium (81.7%) was significantly greater than that of 25(OH) D (75.3%), CA 15.3 (70.1%) and phosphorus (62.8%). The odd's ratio of vitamin D was 0.0937 (95% CI=0.0311-0.2823), of ionized calcium was 0.0464 (95% CI=0.015-0.141) and of phosphorus was 2.6801(95% CI=1.1269-6.3742) in breast cancer patients group. Serum phosphorus was significantly correlated with age and menopausal status of breast cancer patients. Conclusion: Our results suggest that serum ionized calcium and 25(OH) D were superior to serum CA15.3 and phosphorus for prediction of breast cancer. In addition, our results indicate that 25 (OH) D and calcium may decrease the risk for breast cancer incidence, while phosphorus may increase this risk. None of the assayed biomarkers has a prognostic role in breast cancer.