Importance of Early Diagnosis of Invasive Aspergillosis Based on Microbial Cell-free DNA Derived from Invasive Aspergillus Species

Invasive aspergillosis (IA) is a life-threatening opportunistic fungal infection that primarily affects immunocompromised or critically ill patients, and early diagnosis remains a major clinical challenge.Although conventional diagnostic methods are widely used, their clinical utility is often limited by delayed positivity, suboptimal sensitivity during early infection, and the need for invasive sampling procedures.Microbial cell-free DNA (mcfDNA)-based diagnostics have recently emerged as a promising non-invasive approach for the early detection of invasive fungal infections through blood-based molecular assays.By enabling pathogen detection independent of culture or antigen release, mcfDNA analysis offers potential advantages in terms of diagnostic accuracy, earlier detection, and clinical applicability in high-risk populations.Recent studies have demonstrated that mcfDNA-based assays can detect fungal DNA at low concentrations, suggesting their potential utility for early-stage diagnosis and disease monitoring.This review summarizes current advances in mcfDNA-based diagnostic technologies, including polymerase chain reaction (PCR), droplet digital PCR, and metagenomic next-generation sequencing, and discusses their diagnostic performance, clinical applications, and limitations in the context of IA.We also highlight the potential role of mcfDNA-based assays as complementary tools within future diagnostic algorithms for IA.