The aim of the present study was to improve the conventional wire-guided localization biopsy (WGLB) of breast microcalcifications to overcome disadvantages associated with the procedure, including inaccurate localization and large specimen volume. The novel approach described in the present study was termed double wire‑guided localization and rotary cutting biopsy (DWGLB). Prior to surgery, the precise localization of the lesions was assessed using two wires under the assistance of mammography X‑ray and ultrasound, followed by complete excision of the lesions using a novel rotary cutting tool. The cylindrical specimen was placed on a scaled specimen holder for pathological examination. DWGLB was performed in 108 patients with the classification of as Breast Imaging Reporting and Data System score 4A. Percutaneous localization of the lesions guided by a mammography X‑ray and ultrasound were successful in all 108 lesions (100%) with one puncture attempt. The lesions were precisely excised in all of 108 patients, and included 13 malignant lesions (DCIS of breast in 7 cases, DCIS with focal invasive carcinoma in 3 cases and invasive ductal carcinoma in 3 cases). The average distance of the BARD Dualok to the lesion was 4.1 mm; the average weight of specimens was 8.5 g. Compared with WGLB, DWGLB offers several advantages, including more accurate localization of lesions, a more standardized biopsy method and a smaller specimen volume. DWGLB can also provide the precise position of lesions in the specimen for further pathological examination.