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BackgroundTuberculosis, caused by Mycobacterium tuberculosis, is the second leading cause of death from infectious diseases worldwide. Tuberculosis is associated with alterations in sex hormone levels, particularly testosterone, estradiol, and progesterone. However, previous studies have reported conflicting results, with some showing increased or decreased levels in tuberculosis positive patients, while others found no significant differences. This study aims to assess and compare sex hormone profiles among adult tuberculosis-positive patients and tuberculosis-negative individuals and to identify associated factors.MethodA comparative cross-sectional study was conducted from June 15 to August 20, 2024, among 300 eligible adult tuberculosis-positive patients and age-matched tuberculosis-negative individuals in five selected health institutions in Gondar town. Participants were recruited using a simple random sampling technique, and sociodemographic, clinical, anthropometric, and behavioral data were collected through a structured questionnaire. Five milliliters of venous blood were used to determine hormone levels using the Beckman Coulter DXI 800 chemistry hormonal analyzer. Hypogonadism was defined by sex-specific hormones and categorized as primary, secondary, and subclinical. The data were analyzed using SPSS version 25.0. Descriptive statistics, independent t-tests, one-way ANOVA, Mann-Whitney U, Kruskal-Wallis H test, and bivariable and multivariable statistical models were used. A p-value ResultMale tuberculosis-positive patients showed significantly higher estradiol, luteinizing hormone, and FSH (p p p = 0.001), those with sometimes [AOR = 9.2, 95% CI (2.77, 30.62)] (p p = 0.042), and cortisol [AOR = 4.01, 95% CI (1.7, 9.5)] (p = 0.002) levels were significant determinants of hypogonadism in tuberculosis patients.ConclusionMale tuberculosis-positive patients showed significantly higher mean estradiol, luteinizing hormone, and follicle-stimulating hormone but lower testosterone levels, while female tuberculosis-positive patients had significantly lower testosterone and progesterone and higher follicle-stimulating hormone levels. There was a higher prevalence of hypogonadism, particularly in males. Early diagnosis of these hormonal changes is crucial in tuberculosis-positive patients to prevent complications such as impaired gonadal function related to hormonal disturbances.