Mycoplasma gallisepticum (MG) is a worldwide ruined bacteria affecting different avian species, causing severe economic losses. Consequently, the current research sought to detect the incidence of MG among different commercial broiler, layer chickens and turkey farms, and environmental litter samples in different Egyptian governorates (Damietta, Giza, El-Qalyobia, El-Sharqia, and El-Behera) from January 2019 to December 2020. Four hundred samples (infraorbital sinus aspirates, tracheal swabs, serum from diseased birds, and organ samples; lung tissues, air sacs and tracheal bifurcation from freshly dead birds), and environmental samples (litter) were collected for MG isolation. Samples were subjected to phenotypic and molecular identification. Positive bacteriological samples were subjected for molecular identification using polymerase chain reaction (PCR) test to detect MG, then sequencing for PCR amplicon of mgc2 gene. Out of 332 samples subjected for bacteriological examination, 206 were bacteriologically positive for MG with an incidence of 62%. The highest incidence of MG was detected in turkey farms at a rate of 83%, followed by broiler chicken farms, layer chicken farms and litter samples at a percentage of 70, 40, and 40, respectively. The highest prevalence of MG in chickens and turkey was recorded during the winter and autumn seasons. Molecular identification of MG isolates revealed that 85% of isolates were positive for mgc2 gene using PCR. The Four sequenced strains in this study are closely related and placed in one group with the vaccine strain 6/85 and ts11 strain.