A total of 110 chicken samples were collected randomly from naturally infected farms in Al-Rahmania district in Behera Province, Egypt where birds suffered from mottled liver with yellowish discoloration, distended gallbladder, slight air sacculitis, congested lungs and enlarged spleens. Liver, heart, spleen and cloacal swabs from chickens were sampled for isolation of Salmonella enterica serovar Enteritidis (SE). In addition, 50 human stool samples were collected from persons suffering from pyrexial illness with diarrhea and vomiting for several days hospitalized in areas surrounding examined poultry farms to be investigated for presence of SE by bacteriological examination. The obtained results of bacteriological examination revealed that Salmonella spp. were isolated from 4 out of 110 (3.63 %) of examined broiler chicken samples and 2 out of 50 (4 %) of examined stool samples. The serological identification of Salmonella isolates by using polyvalent and monovalent (O) and (H) Salmonella antisera clarified that 3 isolates from chicken isolates were proved to be SE and one isolate from human isolates was proved to be SE. At the same time, all positive samples were re-examined by using Polymerase chain reaction (PCR) where its result revealed that 4 isolates out of 6 were SE. The obtained results of the current work confirmed the diagnostic value of PCR detection ofSE in poultry and surrounding people in which PCR is very sensitive, very specific and relatively rapid test.