Background: In hemodialysis patients, tunneled, cuffed, double-lumen catheters are frequently used for long-term venous access. Infections associated with these catheters are one of the major causes of morbidity and resource usage in the hemodialysis population. Objective: The aim of the current study is to investigate the frequency and types of fungal infection in patients on hemodialysis with double lumen catheter in Imamein Kadhimein Teaching Hospital and Al-Karama Teaching Hospital, Iraq. Material and methods: For the purpose of detecting a fungal infection, 19 blood samples from hemodialysis patients with double lumen catheters were subjected to both BacT/ALERT system (Blood Culture) and molecular approaches (Multiplex Real Time PCR). Results: By using traditional techniques, the fungi Candida spp. (2%), and Cryptococcus spp. (1%) were detected. The following fungi were identified using molecular techniques: (2%) R.oryzae. Conclusion: The nephrologist should be aware of the many fungal infections that patients receiving hemodialysis with double lumen catheters may acquire, including R.oryzae, Candida spp < /em>., and Cryptococcus spp < /em>. The fungi mentioned above may be acquired during hemodialysis or may already be present as underlying illnesses. This study established that hemodialysis patients with renal impairment who use double lumen catheters could have several complications.