This study characterized oropharyngeal fungal colonization in Iraqi transfusion-dependent thalassemia patients. Iron overload poses a risk of infection, but little is known about the local fungal epidemiology. We identified species biochemically and assessed amphotericin B and nystatin susceptibility from clinical isolates. Microbial growth was observed in fourteen of these samples. Following initial culture on Sabouraud dextrose agar, all isolates were sub-cultured on chromogenic agar (CHROMagar™ Candida) to differentiate Candida species based on colony color. Yeasts are recognized as prevailing etiological agents of life-threatening invasive fungal infections in severely immunocompromised thalassemia patients, often necessitating extended intensive care. C. albicans (11 colonies; 72.5%), C. parapsilosis (4 colonies; 22.4%), and C. tropicalis (2 colonies; 5.1%) made up the 17 speciated isolates. Critical virulence determinants, including biofilm formation, germ tube induction, and extracellular phospholipase activity, were profiled in order to assess the pathogenic potential of clinical Candida isolates. In order to perform phenotypic identification, samples were first cultured on Sabouraud dextrose agar (SDA) and then subcultured on chromogenic medium (CHROMagarTM). The results demonstrate that C. albicans isolates were proficient producers of biofilms, phospholipase, and germ tubes, expressing the full complement of virulence factors assayed. This study evaluated the in vitro antifungal efficacy of Nystatin and Amphotericin B against clinical isolates of Candida spp. using the disk diffusion method. Amphotericin B demonstrated superior antifungal activity compared to Nystatin. The speciated isolates (n=17), identified on CHROMagar™ medium, comprised C. albicans (69.9%, n=11), C. parapsilosis (20.5%, n=4), and C. tropicalis (9.5%, n=2). Furthermore, the virulence potential of the predominant C. albicans isolates was assessed, confirming their ability to produce key pathogenic factors germ tubes, biofilms, and extracellular phospholipase enzymes when cultured on specific indicator media.