CONTEXT: Aspergillus is present ubiquitously in the environment in the form of spores. Infection due to Aspergilli is uncommon in immunocompetent individuals, unless they possess any abnormalities or have undergone any treatment with corticosteroids in which pulmonary aspergillosis is the most common. In immunocompromised individuals, the infection by this fungus is in a higher extent of risk. AIM: A retrospective study was done and correlated with the antifungal susceptibility pattern of Aspergillus species. MATERIALS AND METHODS: A total of 72 isolates of Aspergillus were collected and confirmed by conventional methods except Aspergillus tetrazonus was identified by DNA sequencing. Antifungal susceptibility testing (AFST) was performed by conventional broth microdilution according to the Clinical Laboratory Standards Institute M38-A2 2008 to all the isolates. RESULTS: In our study, Aspergillus flavus was found to be the predominant followed by Aspergillus niger being the second most to be identified. AFST was performed for all the isolates, where they exhibited minimum inhibitory concentration (MIC) values within the range of 0.0078–2 μg/ml except Aspergillus terreus (4 μg/ml), which exhibited intrinsic resistance to Amphotericin B. CONCLUSION: Aspergillus infection can be treated easily, unless they become invasive. Therefore, antifungal therapy should be started early by determining the MIC values for the isolates, especially for the systemic infections and prevent from mortality.