Objectives: One of the most significant risk groups for hearing loss is neonates who spend time in the neonatal intensive care unit (NICU). This study aimed to estimate the prevalence of hearing loss in all neonates admitted to our hospital NICU and evaluate their risk factors. It is a prospective cross-sectional study conducted on all neonates admitted to our hospital in one year. Patients and methods: All Neonates were subjected to history, clinical, Otoscopic examination, and Oto-acoustic emissions test. Neonates with congenital ear anomalies or ear problems that interfere with the screening test were excluded. Results: 90.6% of cases passed the test, and 9.4% had a Refer result in the initial screening. A repeated test for the referral cases showed 6.7% passed and 2.7% had hearing affection (2% bilateral and 0.7 % unilateral). Conclusions: The prevalence of hearing loss was 2.7%, 2 % had bilateral hearing affection, and 0.7 % had unilateral hearing affection. There was a significant correlation between exposures to certain risk factors among the affected neonates; all cases remained more than 5 days in NICU with history of exposure to ototoxic drugs, especially gentamycin, furosemide, vancomycin, and amikacin.