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

Abstract

Background Fungi are common opportunistic pathogens that pose a significant threat to immunocompromised patients, particularly when late detection occurs. Methods 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. Results The specificity of the assay was rigorously tested on 40 types of organisms (n = 65), demonstrating 100% specificity. The limit of detection was determined to be 100 pg/μl (106 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. Conclusion This multiplex real-time PCR assay holds promise for enhancing the timely and accurate diagnosis of invasive mycoses, particularly in immunocompromised patient populations.

Keywords

• Multiplex real-time PCR
• Invasive mycoses
• Fungal pathogens
• Specificity