Backgraund: The aim of this study is identifying the most representative pathogens causing infections of the posterior segment of the eye and verifying the correlation between clinical diagnosis and laboratory diagnosis. Moreover, another aim is the verification of the role of Epstein Barr virus that is often in association with other pathogens. Methods and findings: In this study 35 cases of patients with suspected viral infections of the posterior segment of eye are presented. Among these, 18 were found positive for a microorganism with 22 selected positive plating test, 1 patient was tested 2 times and in 3 patients were found more than a virus. The analysis was performed in polymerase chain reaction real time. Analyzed viruses, all of the Herpesviridae family, are: Varicella Zooster ( VZ ) 7, Herpes Simplex ½ ( HSV) 4, Epstein Bar virus (EBV) 5 , Citomegalovirus (CMV) 2. In addition 4 Toxoplasma gondii (TG) were found. Epstein Barr Virus was found 2 times in association with Toxoplasma gondii, once with Varicella Zooster. Conclusions: The pathogenic role of EBV remains uncertain. There is a correlation between the clinical diagnosis and virus isolated. The laboratory diagnosis is important for choice of therapy.