Avian Influenza Virus (AIV) represents a dramatic pathogen among chicken flocks. The current study was carried out for isolation, identification and prevalence of LPAIVH9N2 from chickens and ducks in Damietta province. One hundred and twenty three (123) samples were collected randomly from broiler chickens in Damietta province in Egypt during 2014 – 2018.The samples were prepared, inoculated into ECE at age 9-11day without marked embryonic changes and even rapid slide HA. The samples were testing by real time RT-PCR that revealed 23 positive cases for AIV subtypes H5 and H9 with percentage of 8% and 10,5 % respectively .Analysis of the sequencing data of the (HA) revealed AIV H9N2 subtype in two samples had the mutation represented by (T3I, S5P, I9M, S11N, M58K and T143S) and (T3I, S5P, and S11N) in first and second sample respectively which is considered specific marker of influenza virus mutation. The Phylogenetic analysis of the HA gene showed that our two Egyptian isolates of H9N2 were resemble that isolated from Egypt in 2016 the G1-like lineage. Effective prevention is depend upon the insertion of such strains in any newly prepared vaccines and to construct a strategy for control the risk associated with spread of influenza caused by such virus.