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Invasive pulmonary aspergillosis still constitutes a diagnostic challenge for clinicians due to its non-specific symptoms. HIV infection represents one of the major risk factors due to the progressive failure of the immune system. In this study, Aspergillus serum galactomannan enzyme linked immunosorbent assay was used for the diagnosis of invasive pulmonary aspergillosis in patients living with HIV/AIDS attending University of Ilorin Teaching Hospital Antiretroviral Therapy Clinic, Nigeria. Blood samples were collected from 97 patients affected by HIV/AIDS. Age range of the study population was between 0 and 79 years. Length of the study: 3 weeks. Three out of the 97 samples were positive for the galactomannan assay with an optical density index of 1.1, 0.9 and 1.1 using the 0.5 cut-off optical density value; the concentration level of galactomannan antigen was therefore 2.20 μg/L, 1 μg/L and 2.19 μg/L, respectively. A prevalence of 3.1 % (3/97) was detected; all positive participants have been on ART for more than 3 years (P<0.0001), and 2 out of the 3 had CD4 count higher than 200 cells/mm3 (P<0.0001). Serum Aspergillus galactomannan assay is less invasive for patients and diagnosis can be made in a relatively shorter time compared to other techniques used in diagnosing invasive pulmonary aspergillosis.