Abstract: The present invention describes a novel molecular method based on the polymerase chain reaction (PCR) in a multiplex variant in order to detect and identify Candida species with clinical relevance, namely C. albicans, C. glabrata, C. krusei, C. parapsilosis, C. tropicalis, C. guilliermondii, C. lusitaniae e C. dubliniensis. The strategy uses the existence of sequences, whether conserved or variable, in fungal ribosomal genes and in the use of a combination of universal primers, specific for fungi, and internal primers, specific for each one of the Candida species (FIG. 1). In this sense, two fragments from the internal transcribed spacer (ITS) regions of the ribosomal RNA (rRNA) are amplified by multiplex PCR, allowing the easy identification of the Candida species in question. This methodology allows a rapid, effective and low-cost identification of Candida species with clinical relevance, bearing several advantages over the currently available diagnostic methods.