Background: Candida dubliniensis that was first identified as a new species by Sullivan et al. (1995) in Dublin, Ireland (and was subsequently named after its place of origin) while performing an epidemiological investigation of oral candidiasis in HIV-infected and AIDS patients in the early 1990s. This pathogenic Candida species shares many phenotypic features with Candida albicans which cause problems its identification. Several phenotypic based tests have been developed to distinguish C. albicans from C. dubliniensis but none has been demonstrated being sufficient alone for accurate differentiation of the two species. Aim: To facilitate the differentiation of these species, we evaluated methyl blue Sabouraud dextrose agar medium. Method: Two hundred Candida spp. were tested including 186 stock strains of C. albicans and 14 strains of C. dubliniensis. Identification of all these strains was confirmed by polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) using BlnI (AvrII) enzyme. All isolate were inoculated on the medium, incubated at 37 __ampersandsigndeg;C in ambient air for 24 to 96h. Examination was done in Fluorescent chamber with illumination at 365 nm. Result: On this medium, 156 C. albicans isolates showed fluorescence at 48h of incubation while none of the 14 C. dubliniensis isolates did so even on extending the incubation period. Also after 96h of incubation colonies of all 14 test strains and the two reference strains of C. dubliniensis showed yellow colour when viewed against light while others did not. Conclusion: In conclusion, based the results of our study, methyl blue SDA test offers an additional simple means for identification of C. dubliniensis.