Mycoplasma gallisepticum (MG) is an important avian pathogen causes significant economic losses within the poultry industry. One of the options of the controlling MG infection is the isolation and identification of MG.215 samples collected from the lungs, tracheal biforcation, tracheal swabs, blood samples and frontal sinus aspirates were collected from broiler and layer flocks in Damietta governorate. Samples were cultivated on Frey's media giving 142 samples positive as (F. Mycoplasmataceae) as 66%. The (142) samples was used for digitonin sensitivity test and the results was (98) samples as digitonin positive (Mycoplasmataceae) (69%) and (44) samples was digitonin negative (Acholeplasmataceae) (31%).The (215) samples was characterized by PCR by using MG primer