Mycoplasmosis in turkeys is a major egg transmitted disease characterized by infra-orbital sinusitis, reduced hatchability, high embryonic mortality, reduced live body weight. Swollen infra-orbital sinus in turkeys resulting in misshapping, and difficult marketing as well as spread of infection to other poultry species.  So, this study aimed to investigate Mycoplasmas involved in turkey sinusitis using molecular technique, their phylogenetic analysis versus GenBank database. Suspected turkey flocks (n=17) were investigated using PCR technique for aspirated fluid from infra-orbital sinus. Seven selected isolates of both identified M. gallisepticum (MG), and M. meleagridis (MM) were subjected for sequencing, and phylogenetic analysis. PCR results exhibited single infection of MG and MM in 8, and 3 out of 17 turkey flocks respectively, while mixed infection of both MG and MM were found in 4 flocks. Sequenced MG mgc2 gene of selected seven strains showed similar identity of 93.1-97.6 with each other, 90.4-97.3% with field strains and 84.9-95.9% with reference ones.  On the other hand, sequenced 16s rRNA of all isolated seven MM strains revealed complete identity of 100% with each other, 99.7% with MM reference strains, and  83.1-93.1% with other Mycoplasmas spp  Both MG, and MM isolates were registered on the GenBank with accession numbers; MW713803, MW713804, MW713805, MW713806, MW713807, MW713808, MW713809, MW700292, MW700293, MW700294, MW700295, MW700296, MW700297, MW700298. It was concluded that the increased incidence of MG in turkeys sinusitis, also, MM isolates were the 1st Egyptian field strains submitted to GenBank database. There were similarities between the recent isolates and reference vaccine strains as well as these molecular analysis and genetic diversity would play an important role in control strategy of Mycoplasmosis in poultry.