Abstract: Objective：To analyze the false positive rate and false negative rate of layered mammography in diagnosis of mammary gland diseases. Methods：120 patients with malignant breast diseases and 120 patients with benign breast diseases treated in Huadong Hospital Affiliated to Fudan University from February 2015 to October 2016 were selected. All patients were examined by both traditional mammography and layered mammography. Taking surgical pathology as the gold standard, the diagnostic value of the two methods on patients with different ages, different lesion sizes， and different gland texture were compared. Results：The false positive rate and false negative rate of patients with mammary gland diseases in layered mammography group aged less than 45 years, 45-60 years， and more than 60 years were lower than those of the conventional group, and the differences were statistically significant (P<0.05). The false positive rate and false negative rate of patients with mammary gland diseases in layered mammography group with lesion sizes less than 1 cm, 1-2 cm, and more than 2 cm were significantly lower than those of the conventional group, and the differences were statistically significant (P<0.05). As for the patients with different gland texture, the false positive rate and false negative rate of the two groups were not statistically different. Conclusions：The false positive rate and false negative rate of layered mammography in diagnosis of breast diseases are lower than the conventional mammography, so it has higher clinical diagnostic value.