Bckground/aim: Invasive aspergillosis (IA) is a fatal infection that is difficult to diagnose in immunocompromised patients. In thisstudy, Aspergillus-specific DNA was searched using real-time PCR (RT-PCR) in serum samples. Galactomannan (GM) and/or beta-Dglucan (BDG) tests were previously performed on these samples for 70 neutropenic patients with hematological malignancy.Materials and methods: The patients were categorized according to the criteria of the European Organization for Research andTreatment of Cancer/Mycoses Study Group (EORTC/MSG). Among the patient serum samples, the first positive GM or BDG testsample and the median sample of GM or BDG test for negative patients were used to detect DNA levels by RT-PCR method (LightCycler 480, Roche Molecular Biochemicals, Meylan, France) using a commercial kit (Way2Gene Fungi; Genmar, İzmir, Turkey).Results: When the proven and probable IA group were considered as real patients, sensitivity of Aspergillus-specific DNA test was 90%,specificity was 73.3%, positive predictive value was 81.8%, and negative predictive value was 84.6%.Conclusion: This study found that searching for specific DNA by RT-PCR method has a sensitivity as high as the GM test. Althoughspecificity was rather low, it was concluded that it can be used jointly with GM and BDG tests after decreasing contamination by severelaboratory applications.