To evaluate the strain ratio (SR) combined with molecular pathological and serum markers for the diagnosis of breast masses. SR and 7-point scale elasticity scores were used with real-time tissue elastography and 2-dimensional color-Doppler ultrasound (US) to diagnose breast lesions in 311 hospitalized patients. Immunohistochemical staining and enzyme-linked immunosorbent assays (ELISAs) were used to examine pathological and serum tumor markers and their correlations with SR findings. SR had a higher diagnostic value compared to the 7-point scale elasticity score, displaying an obvious low-to-high distribution from benign to malignant lesions with an optimal cutoff point at 3.88, which yielded an area under the curve (AUC) of 0.896 with 89.1% sensitivity, 85.6% specificity, and positive and negative predictive values of 91.0% and 82.8%, respectively. The differences of SR values between small (≤1.5 cm), large (>3 cm) (P=0.010), and moderate (>1.5 cm and ≤3 cm) sizes (P=0.038) in distinguishing benign from malignant breast masses were statistically significant, with SR being most specific and sensitive for diagnosing small lesions. Expression of 3 molecular pathological indicators (p75NTR, p63, and CK5/6), and 5 serum mastocarcinoma markers (uPA, PAI-I, CA27-29, CEA, and CA15-3) showed statistical significance (P<0.05) in distinguishing between benign and malignant breast lesions. Furthermore, SR combined with CA15-3 and CK5/6 positivity showed 94.2% sensitivity and 89.2% specificity as combined markers for triple-negative (TN) breast cancer, whereas SR combined with D2-40 and CK19 were good diagnostic markers for breast cancer lymph node metastasis. SR, together with a molecular and serological marker, may serve as an additional tool for the diagnosis of small breast cancer tumors.