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Objective: Treatment with aminoglycosides is a preventable cause of acute kidney injury (AKI). Early predictive markers of AKI are crucial in preventing this condition. This study aimed to examine the role of serum neutrophil gelatinase-associated lipocalin (NGAL) level as a predictive marker of AKI in patients receiving gentamicin. Materials and Methods: In this prospective study, 37 patients with brucellosis (23 males, 14 females; mean age = 10.7 ± 33.7 years) were studied in a health center between March 2013 and March 2014. Serum creatinine and NGAL levels at baseline, and 3, 5 and 7 days after the administration of gentamicin were measured. In this study, a 25% increase in the serum creatinine levels compared with the baseline levels was considered a criterion for AKI risk. Results: The frequencies of patients prone to AKI at the first, third, fifth and seventh day of administration of gentamicin were 0%, 8.1%, 18.9%, and 13.5%, respectively. In total, 24.3% of the patients (n = 9) were prone to AKI. No statistically significant difference was observed between NGAL levels before and after the administration of gentamicin (P = .082). Conclusion: The serum NGAL level is not a sensitive and specific predictor of AKI following the administration of gentamicin. Therefore it is recommended to increase the frequency of sNGAL measurement and using more sensitive kits in more patients.