BACKGROUND: Tuberculous is the leading cause of death worldwide with a large number of deaths occurring in developing countries. Tuberculous lymphadenitis is among the most common presentations of extra pulmonary tuberculous. This study attempts to determine the magnitude of tuberculous lymphadenitis from patients with lymph node aspirate in Gondar University Hospital, Northwest Ethiopia. METHODS: Retrospective study was conducted. Data were collected from registration book of Gondar university Hospital pathology laboratory after checking the completeness of patient’s necessary information like age, sex and fine needle aspiration cytology results. Data were entered and analyzed using SPSS version 16 statistical package. Chi-square test was done to determine associations. RESULT: A total of 3,440 lymph node aspirates were examined using fine needle aspiration cytology. Of these, 2,392 (69.5%) cases were found to have tuberculous lymphadenitis. Male 1647(47.9%) to female 1793(52.1%) ratio of all study subjects were 0.9:1. Females (54.1%) were more affected than males (45.9%). Age, sex and site of aspiration were found to be statistically associated with tuberculous lymphadenitis (p-value < 0.001). The age group of 15-24 years had the highest prevalence of tuberculous lymphadenitis followed by those of 25-34 years old. The most affected sites were cervical lymph nodes (47.5%) followed by auxiliary (19.4%) and submandibular (12.9%) lymph node regions. None of the records documented the HIV status of subjects. CONCLUSION: The prevalence of tuberculous from lymph node aspirate was found to be higher involving the frequently affected site of cervical lymph node. The HIV status of patients with all forms of tuberculous should have to be checked and documented. Further prospective and advanced studies are recommended to determine the specific etiologic agents and contributing factors.