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Invasive Fungal Infections (IFIs) are a growing global health concern, particularly among immunocompromised individuals and critically ill patients. Diagnosis remains challenging due to nonspecific symptoms, low sensitivity of conventional methods, and the emergence of antifungal resistance. This review outlines the diagnostic limitations of microscopy, culture, and serological assays while exploring the potential of molecular tools. Emphasis is placed on integrated diagnostic pathways, resistance gene detection, point-of-care assays, and Artificial Intelligence (AI) enhanced imaging for fungal infections. Diagnostic challenges in special populations are also addressed. The role of diagnostics in antifungal stewardship and infection control is discussed with forward-looking perspectives. By integrating conventional approaches with emerging technologies, this review highlights the need for a more precise and responsive diagnostic era in IFIs. Timely and accurate diagnosis of IFIs is vital for improving outcomes. The integration of traditional and emerging diagnostic tools, including pan-fungal platforms and resistance profiling, is key to advancing fungal disease management globally, particularly in resource-limited settings.
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