Real-Time PCR assay for detection of candida parap-silosis and candida orthopsilosis

Abstract

C. parapsilosis and C. orthopsilosis emerged as fungal pathogen with significant worldwide prevalence, particularly in causing nosocomial and skin infections. In this study, we aimed to develop molecular assay based on real-time PCR for sensitive and accurate detection of C. parapsilosis and C. orthopsilosis. A pair of primers that specifically target on both of these yeast species was designed and real-time PCR amplification assay was optimized using EvaGreen as the DNA binding dye. The optimized assay could detect and quantify up to 1 pg concentration of C. parapsilosis and C. orthopsilosis DNA with amplification efficiency of 104% and 103%, respectively. Both the designed primers and the quantitative assay will have a great potential as molecular diagnosis tool for early detection of fungal infection caused by either C. parapsilosis or C. orthopsilosis, which merits future clinical study prior to use in diagnosis.