Molecular Identification of Trichosporon Asahii Used DNA Sequencing Methods Isolated from Patient with Diabetic Foot

Abstract

The aim of this study was conduct and investigate to the genotyes identification ofT. asahii isolates obtained from patients with diabetic foot in Al-Najaf province-Iraq , One pathogenic yeast, T. asahii, was used in this study isolated from patient with diabetic foot The first step they were activated all isolates on Brain heart infusion broth for 2 days at 25°C and examined for purity ,second step involved morphological and microscopic examination, to verify the isolation's identity by staining with lactophenol stain, DNA extraction and purification using Accu power PCR Pre Mix kit with both (ITS1-ITS4) primers ,The PCR product was then sequenced using the Sanger method by delivered to Macrogen Lab in the United States .The colonies were usually raised and have a waxy appearance, also developed furrows and irregular folds, on SDA after 3 days at 37°C, T. asahi that was discovered on chromagar, who appeared pale lavender with a white rim of T. asahi on chrome agar plate after 2 days of incubation at 37°C .The results of primer pair which was targeted ITS regions sample 30 refered to T. asahi with moleculare weight 736 pb .The strain L3_LF was found to be the nearest neighbor to T. asahii strain KTSMBNL-12 with identity 99.21% while the strain L3_LR was found to be the nearest neighbor to T. asahii strain AUMC 10759 with identity 98.97% .