Since Mycoplasma Gallisepticum (MG) is one of the most pathogenic Mycoplasma, and caused tremendous economic loss in the poultry industry. Therefore, the present study aimed to isolate MG from chicken tracheas samples and to confirm molecular identification the MG strain using PCR. Mycoplasma gallisepticum which isolated was identified by the polymerase chain reaction (PCR). Banding patterns that were clearly identical from the common MG Ts-11 strain and in MG isolated from the field at 888-938 bp. Our data demonstrated an excellent technique to diagnose and investigate MG infections in chickens. The isolated MG can be used as autogenous vaccines in the future.