Recurrent vulvovaginal candidosis is a frequent disease with a serious impact on women's quality of life. Mostly, recurrences are caused by identical Candida strains suggesting C. albicans persistence in the female anogenital area. Objectives of the presented work were to identify the site of C. albicans persistence, to determine clinical symptoms and signs related to C. albicans positive vulvar cultures and to introduce a new therapeutic approach in women with RVVC. Women with an acute, culture-confirmed episode of RVVC at time of visit were included in this prospective case series. Swabs were obtained from both vagina and inter-labial sulcus. Women received a combined 20-day regimen of 100 mg oral fluconazole and ciclopiroxolamin cream topically. Follow-up visits were at 3, 6, 9 and 12 months. Of 139 women, 105 (76%) had at least one C. albicans positive culture from the external vulva. Vulvar positive cultures correlated with pruritus (OR 5.4; P < 0.001), vulvar edema (OR 3.8; P = 0.03) and fissures (OR 2.4; P = 0.03). Recurrence rates were 27%, 33% and 34% (at 6, 9, 12 months, respectively). The external vulva appears to represent a site of C. albicans persistence and source of endogenous re-infection in patients with RVVC. The combined treatment compared favorably with published fluconazole maintenance regimens.