Background: Candidiasis is one of the prevalent fungal infections caused by the genus Candida. The clinical manifestation ranges from mucocutaneous colonization to disseminated and fatal infections such as candidemia. Diabetes mellitus is one of the significant predisposing factors for this fungal infection. Candida spp. may release many exoenzymes such as phospholipase to debilitate the immune system and facilitate adherence and invasion of the fungus to the host cells. The aim of the present study is evaluation of phospholipase activity of Candida species isolated from candidemia and gastroesophageal candidiasis (GEC) among diabetic patients. Materials and Methods: Eighty-three Candida isolates were evaluated for enzyme activity by phenotypic (the precipitation zone around the colonies) and molecular methods (detection of phospholipase genes using duplex polymerase chain reaction with specific primers). Results: Eight out of eighty-three clinical isolates (9.6%) were negative for phospholipase production. All phospholipase producers among candidemia and GEC isolates were categorized in high production group. Conclusions: Our findings revealed no differences in phospholipase activity among isolates obtained from different body sites (blood, oesophagus and stomach); however, non-albicans Candida species had less phospholipase activity.