Simplex and triplex polymerase chain reaction (PCR) for identification of three medically important Candida species

Abstract

Candida species are a major cause of invasive infections in both critically ill and immunocompromised patients. Hence, rapid identification of these pathogens may facilitate specific therapy and patient management. The development of rapid and specific diagnostic methods remains a challenge. Herein, we developed the simplex and triplex polymerase chain reaction (PCR) for the identification of three medically important Candida species namely C. albicans, C. parapsilosis and C. tropicalis. The developed methods target the phospholipase B gene (PLB). The primers designed achieved highly specific identification of the selected species using both the simplex PCR and the triplex PCR formats, which were confirmed by DNA sequencing. The primers did not show any non-specific amplification when tested with DNA from other Candida species and other fungal species such as Aspergillus and Cryptococcus. These results showed that the PLB gene provides a novel target that could be used for the detection of medically important Candida species from clinical specimens.

Key words: Candida species, primers, phospholipase B gene (PLB), polymerase chain reaction (PCR).