The applicability of luminescent immunoassay (LIA) in serodiagnosis of fungal infections in multitransfused (MT) thalassemic children seropositive for human immunodeficiency virus (HIV) was investigated. Thirty-one sera samples from HIV infected pediatric patients with thalassemia receiving repeated blood transfusions were analysed for the presence of antibodies specific to Aspergillus fumigatus by LIA. The LIA was standardized using well defined antigens of A. fumigatus. Ten out of 31 (32.2%) of the MT-HIV positive patients were found to have anti-Aspergillus antibodies in their sera by LIA. The ELISA could detect A. fumigatus specific antibodies in 25.8% (8 out of 31) of the patients. Thus, 20% more number of patients turned to be positive for aspergillosis by LIA as compared to ELISA. The difference was found to be statistically significant (p < 0.005). Of the MT-HIV negative patients only 1 out of 33 (3%) showed A. fumigatus specific antibodies by LIA and ELISA both. In age and sex matched control group (n = 25) none of the patients was found to be positive for antibodies to A. fumigatus. LIA was found to have better discriminatory value indicating, thereby, its utility in diagnosis of aspergillosis in compromised patients.