The frequency of fungal infections, especially of Invasive Fungal Infections (IFIs), has risen dramatically in recent years, and has high morbidity and mortality. Early and accurate diagnosis is important for timely implementation of antifungal therapy and decreasing the unnecessary use of toxic antifungal agents. Conventional microbiological, histological and radiological techniques remain the cornerstone of diagnosis but are insensitive and have a limited impact on clinical decision-making. There is always a need to characterize fungi quickly and accurately which can be done by rapid diagnostic techniques like antigen detection and molecular techniques. This is a brief review of all the microbiological techniques-conventional and recent alternative techniques especially elaborating antigen detection and molecular techniques which are available for diagnosis of fungal infections.