Abstract: Teliospores of Ustilago scitaminea were collected from whips of infected sugarcanes and cultured on YGC medium which conducted to isolation of yeast-like and mycelial colonies. White mycelia were also isolated from culturing of pieces of infected tissues of sugarcane. Results of PCR using specific primers bE4 and bE8 revealed the fungal detection in extracted DNA from yeast-like and mycelial colonies. PCR reaction was specifically amplified a DNA fragment of 495 bp in all three types of colonies of yeast-like and mycelial forms. This DNA fragment was not amplified in DNA extraction of yeast-bacterial colonies and other saprophytic-mycelial fungi.