Background : Human Adenovirus (HAdV ) is considered one of the important virus that infect humans and cause multiple infections in the body,it is one of the important causes of conjunctivitis , respiratory and gastrointestinal infections, causing diarrhea. The current study was done to determine the genotype of HAdV in Egypt. Materials : The study was conducted from November 2021 to November 2022. 120 samples ( 90 stool samples and 15 conjunctival and 15 respiratory samples ) were collected from different regions of Egypt, Information related to age, sex and region were recorded . The HAdV was detected serological by ILFST test . The positive samples were cultured and HAdV isolated on Vero cell lines . HAdV was collected from infected cells and then identified using the multiplex PCR technique . The genotype HAdV was determined through bioinformatics. Results : Through this study , ILFST test , HAdV was detected in 20 (22.22%) out of 90 stool samples ,while the conjunctival and respiratory samples was negative. 12 out of 20 samples were cultured on Vero cells . The cytopathic effect CPE was observed with 6 out of 12 samples . Three samples were identified as adenovirus using the multiplex PCR , and confirmed by agarose with expected 350 bp .