Objective To compare and evaluate the efficacy of diagnosis and excision for appropriately selected breast multi-focal lesions and solitary lesion by ultrasound-guided vacuum-assisted biopsy (UGVAB). Methods Among 392 appropriately selected patients, 187 patients with multi-focal lesions and 205 patients with solitary lesion were treated by the 8-gauge UGVAB from May 2007 to June 2009. All lesions were removed as completely as possible. The patients with benign pathology underwent physical and ultrasound examinations at one week and 6 months after procedure. Results During the procedure, only three patients had vasovagal syncope and twenty others complained of other intraoperative discomfort. An accurate pathological diagnosis was obtained in all lesions. There was no apparent false-negative result among the 696 lesions with benign pathology at a follow-up of 6 months after procedure. The rates of malignant or premalignant pathology, postoperative complications and residual lesions in patients with multi-focal lesions were higher than those in patients with solitary lesion. If each lesion was considered as a subject of study, there was no significant difference between the two groups. Conclusion UGVAB is an effective method for diagnosis and excision of appropriately selected breast multi-focal lesions and can be used routinely.