In the present retrospective study, the clinical value of color Doppler ultrasound combined with the detection of the serum tumor markers carbohydrate antigen 153 (CA153), carcinoembryonic antigen (CEA) and tumor specific growth factor (TSGF) as diagnostic tools for breast cancer was investigated. A total of 103 patients with breast cancer and 50 patients with benign breast lesions were enrolled in the study. All patients were examined by color Doppler ultrasonography. Electrochemiluminescence was used to measure the expression levels of CA153 and CEA, and chemical colorimetric assay for the measurement of TSGF expression. The differences in mass boundary, morphology, internal echo, calcification, peak blood flow velocity (V max ), resistance index (RI), pulsatility index (PI) and blood flow signal classification between the two groups were statistically significant (P<0.05). The expression levels of CA153, CEA and TSGF in the serum of patients in the breast cancer group were significantly higher than those in the benign lesion group (P<0.01). In the breast cancer group, CA153, CEA and TSGF expression levels were significantly higher in patients with high-stage cancer and recurrence, compared with the patients with low-stage cancer and no recurrence. The sensitivity, accuracy and negative predictive value of the combined detection were significantly improved compared with those of the single tests (P<0.01). Color Doppler ultrasound combined with the detection of CA153, CEA and TSGF levels in the serum of patients can be used as an effective tool that can improve the accuracy of breast cancer diagnosis leading to early diagnosis and clinical intervention.