Development of multiplex real-time PCR for simultaneous detection of common fungal pathogens in invasive mycoses

Fungi are common opportunistic pathogens that pose a significant threat to immunocompromised patients, particularly when late detection occurs. In this study a multiplex real-time PCR has been developed for simultaneous detection of common fungal pathogens associated with invasive mycoses in a diagnostic setting. The specificity of the assay was rigorously tested on 40 types of organisms ( = 65), demonstrating 100% specificity. The limit of detection was determined to be 100 pg/μl (10 copies/μl), achievable within a rapid 3-h timeframe. The PCR assay efficiency exhibited a range between 89.77% and 104.30% for each target organism, with linearity falling between 0.9780 and 0.9983. This multiplex real-time PCR assay holds promise for enhancing the timely and accurate diagnosis of invasive mycoses, particularly in immunocompromised patient populations.