Conventional methods for diagnosis of invasive aspergillosis (IA) lack sensitivity and specificity. Serological methods still have many cases of cross-reactivity. However, molecular techniques seem to arise as a rapid approach, specific and direct that could be used in the diagnosis of IA. In this study, we analyzed 88 serum samples from patients of having IA using GM-ELISA test, nested PCR with primers for the rRNA 18S of Aspergillus genus and real time PCR specific for A. fumigatus. Among the 88 samples, 64 of them had positive GM titers and 23 had positive nested PCRs; 18 of the 23 PCR-positive samples were also GM-positive. On the other hand, 18 samples were detected positive by reel time PCR; 13 positive samples were also detecting positive by nested PCR. QPCR revealed 26 % of the patients with IA, while nested PCR and galactomannan ELISA revealed respectively 34 % and 94 % of the patients with IA. Probable IA was diagnosed in 18 and possible IA was diagnosed in 6 episodes. Forty-four episodes were defined as not having IA. The positive and negative predictive values were respectively 100 %, and 88 % for QPCR, 100 % and 97 % for nested PCR, and 28 % and 60 % for GM test. These results suggesting that combined use of methods might improve the diagnosis of IA.