Recently, relentless efforts to develop rapid, cost-effective, and reliable laboratory methods for daily diagnosis of fungal diseases such as aspergillosis appear to be materialized in the relatively new, but revolutionary matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry (MS) technology. As for Aspergilli, MALDI-TOF MS profiling of isolates growing in culture--characteristic protein spectra are obtainable by means of simple and reproducible preanalytical and analytical procedures--ensures that single species within the different sections or complexes can be easily and accurately identified, including species that are morphologically and phylogenetically similar to each other. Thus, resort to longer and more onerous molecular biology techniques is restricted to those cases for which no spectra in the reference fungal database or library are available at the time of analysis. However, it is necessary to interrogate reference libraries composed of spectra that have been obtained using procedures similar to those used to obtain the test isolate's mass spectrum, as well as to continuously update these libraries for enriching them with fungal strains/species not (or not well) represented in their current versions. Compared to mold identification, very limited work was reported on the use of MALDI-TOF MS to perform strain typing or antifungal susceptibility testing for Aspergilli. If these complementing areas will be potentiated in the near future, MALDI-TOF MS could effectively support the clinical microbiology/mycology laboratory in its primary role of assisting either infection control specialists or physicians for the diagnosis and treatment of aspergillosis.