Molecular identification of a 65 kDa mannoprotein encoded gene of Candida species by PCR

 Introduction: Invasive fungal infections represent a major public health concern. In particular, systemic candidiasis remains an increasing source of morbidity and mortality especially in immunocompromised patients such as neutropenic patients undergoing antiblastic chemotherapy or bone marrow transplants. Early diagnosis is often difficult. Consequently, effective treatment is often delayed. PCR has the potential to decrease the time required to diagnose fungal infections and therefore reduce the mortality associated with disseminated disease.The MP65 gene of Candida albicansis appropriate for detection and identification. The aimofthisstudy was toidentifydifferentspecies of Candida (C. albicans, C. glabrata, C. parapsilosis) withPCRtechnique. Materials and Methods: All 107 yeast isolateswere identified on cornmeal agar supplemented with tween-80, germ tube formation in serum, and assimilation of carbon sources in the API 20 C AUX (Biomerieux, France). Then, all isolates were tesed by PCR using by different species-specific PCR primers selected within the MP65 gene a recently cloned gene encoding a mannoprotein adhesin. Results: A hundred of clinical isolates were detemined as C. albicans and 6 of the isolates detemined asC. glabrata and 1 of the isolates detemined as C. parapsilosis. The species-specific PCR primers allowed differentiation of each of three Candida species by the amplicon length produced.The primers amplified all Candida species DNA tested (100% positivity) giving a band of the expected length (475 bp for C. albicans, 361bp forC. glabrata, 124bp forC. parapsilosis). The results were agreed with all culture results for Candida species detection. Conclusion: The results of this study showed that PCR method for rapididentification ofCandida specieswith specific primers is reproducible, simple and specific