Vaginal candidiasis (VC) is a fungal or yeast infection of the vulva and/or vagina. It causes a smelly, thick, whiteyellow discharge that might be accompanied by itching, burning and swelling. Vaginal candidiasis is the second most frequent infection of the female genital tract. Diabetes mellitus is a chronic, insidious disease that can affect any organ of the body. One of the problems associated with this condition is infection patients with diabetes mellitus are at increased risk of vaginal candidiasis. The purpose of this study was to determine the prevalence and species distribution of Candida species isolated from vaginal candidiasis infection in diabetic and non-diabetic women in Sana’a City. This descriptive – analytic study was performed on 150 diabetic women referred to Al-Thaourah and AL-Gmohore hospitals in Sana’a City. All specimens were diagnosed or examined under direct microscopy and cultured on Sabouraud dextrose agar (SDA) or (YEPDA) media. Complimentary tests such as germ tube test, corn meal agar media, CHROM agar test and sugar fermentation test were carried out to differentiate between the Candida species. Minimum inhibitory concentration (MIC) test was performed for positive cases. Patient’s clinical history information was collected by a questionnaire. Statistical analysis was performed using the Chi-square test. Of 150 samples tested under all biochemical experiments and culture methods, 62 out of 150 with percentage 42% were infected to vaginal candidiasis. The frequencies of the isolated Candida species include Candida albicans 33 species with 53%, Candida glabrata with 19 species with 31%, Candida tropicalis 6 species with 10% and Candida krusei 4 species with 6%. Vaginal candidiasis was more prevalent in women infected with diabetes mellitus than ones with non-diabetes mellitus women. 11patients out of 150 patients with percentage 8% of the patients had type I diabetes mellitus and 51 patients out of 150 with percentage 34% of them type II with ratio 18:82 from totally of 62 out of 150 with 42% vaginal candidiasis patients had infected. C. albicans was, by far, the most predominant yeast isolates. The culture technique of vaginal discharge not be warranted so we used the microscopic and biochemical tests because are more sensitive in identification of Candida species than culture methods. But vaginal culture is indispensable to confirm the diagnosis by microscopic examination.