This case report describes a new treatment of an isolated longitudinal vaginal septum (LVS) by office operative vaginoscopy with a 4-mm rigid hysteroscope in a 27-year-old virgin who reported leukorrhea and recurrent vaginal infections. This technique might represent an effective treatment of an LVS, mostly in patients with an intact hymen. This could allow the inclusion of vaginal lesions among the indications for office endoscopic procedures performed using operative hysteroscopes.