Objective: To investigate the application value of high frequency B-mode ultrasonography in breast cancer screening. Methods: A total of 13 535 women aged 35–69 years in Guilin, of whom 11 167 cases were from urban areas and 2 368 cases were from rural areas, underwent breast cancer screening that included clinical breast examination, high frequency B-mode ultrasonography and selective molybdenum target mammography. Results: Eight cases of breast cancer (malignant breast tumor) were detected in the entire group by final pathological identification, and the detection rate was 0.591‰ (8/13 535). Of the 8 women with breast cancer, all were positive by high frequency ultrasonography, and the sensitivity was 100%; while in molybdenum target mammography, 7 cases were positive and one case was false negative, and the sensitivity was 87.5%. Conclusion: High frequency B-mode ultrasonography has high sensitivity in breast cancer screening. It has the advantages of non-invasiveness, low cost, wide range of applicability and easy manipulation.