The study was performed on 225 chicken (72 diseased and 153 freshly dead ones) from different 45 commercial broiler farms (1- 30 days old) at Dakahlia Governorate for mycotic infection. Samples were taken from these chickens (lung; air sac; crop; liver and brain from each chicken) after clinical and postmortem examination for mycological examination. The results revealed that, fungi were isolated from 503 positive samples (44.7%); represented as 119 positive samples (10.6%) from diseased chickens and 384 positive samples (34.1 %) from freshly dead ones.651 fungal isolates (moulds and yeasts) were isolated from 1125 samples, where 219(33.6%) were isolated from lung samples followed by 199(30.6%); 119(18.3%); 91(14.0%) and 23(3.5%) from air sac, liver, crop and brain samples respectively. Moreover, A. fumigatus was the most isolated one 141(21.7%) followed by A. flavus 126(19.4%); A. niger 111(17.1%); Rhizopus spp. 67(10.3%); C. albicans 58(8.9%); Mucor spp. 42(6.5%); Penicillium spp. 32(4.9%); A. terreus 20(3.1%); Fusarium spp.19 (2.9%); C. krusei 13(2.0%); C. tropicalis 9(1.4%); A. candidus 9(1.4%) and finally A. ochraceus 4(0.6%). PCR using fungus-specific universal primer pairs (ITS1 and ITS4) was used for identification and genotypic characters of Aspergillus species (A. fumigatus; A. flavus and A. niger).They were able to successfully amplify the ITS1–5.8S rDNA region of all tested Aspergillus isolates, providing a single PCR product of about 600 bp for all tested isolates PCR using fungus-specific universal primer pairs (ITS and RPS) was used for identification and genotypic characters of C. providing a single PCR product of about 109 bp .Sequence of the internal transcribed spacer 1 reigon (ITS-1) of isolated A. flavus Gene Bank accession number for studied nucleotide sequence (Bankit 1867303) is KM 983253. The sequences obtained for ITS-1 region were more than 98% identical to the corresponding GenBank sequences (accession no. DQ467968.1; KC994648.1; LN482516.1 and KP689246.1). Sequence of the internal transcribed spacer 1 region (ITS-1) of isolated C. albicans. Gene Bank accession number for studied nucleotide sequence (Bankit 1867955) is KM 983254. The sequences obtained for ITS-1 region obtained were more than 95% identical to the corresponding GenBank (accession no. M87288.1; gb|S71769.1|S71769; emb (AL033396.1) and AP006852.1).