BACKGROUND Adenoid cystic carcinoma (ACC) often occurs in the major and minor salivary glands and other sites containing secretory glands, while ACC of the Bartholin's gland (BG-ACC) in the vulva is rare and easily misdiagnosed. METHODS A 58-year-old female was referred to our hospital for further valuation of a mass occurring on the left side of her vulva. In the other hospital, the beginning of the period, local ultrasound showed a vulva mass, which was suspected to be a Bartholin's gland cyst. Mixed neoplasms were considered in some biopsies. When transferred to our hospital, virtuous tumors were considered by ultrasound and magnetic resonance imaging. Pathology initially considered benign hyperplastic active tumor or borderline tumor. METHODS Histological, immunochemical, and molecular tests confirmed a diagnosis of BG-ACC, negative surgical margin, without lymphatic metastasis. METHODS Extended excision of the mass at left labia majora plus left inguinal lymph node dissection was performed. RESULTS The patient received surgery therapy, no recurrence was observed during a 18-month follow-up period. CONCLUSIONS Due to its lack of specific characteristics in clinical, ultrasound and imaging, it is easy to be misdiagnosed, Due to its rarity and nonspecific clinical, radiologic and ultrasonographic manifestations, BG-ACC can be easily misdiagnosed. And its pathomorphological features overlap with other benign and malignant tumors occurring at vulva, BG-ACC can be easily misdiagnosed, and diagnosis by puncture biopsy is extremely difficult. Use of paraffin sections to identify tumor growth characteristics, combined with immunohistochemical findings, is the key to the diagnosis of ACC. In rare sites, MYB gene split are helpful in making a definite diagnosis.