Aims: Brucellosis is one of the important infectious factors in most areas of Iran and other developing countries. Brucellosis has different clinical manifestations and should be considered in the differential diagnosis of infectious and noninfectious diseases. The aims of this study was to determine the epidemiological, clinical findings of the brucellosis in patients based on blood culture and serological tests. Materials and Methods: In this cross-sectional study, 190 brucellosis patients admitted in Isfahan Hospital in 2016–2017 that were studied based on positive immunological tests such as Wright, 2-mercaptoethanol (2ME), and enzyme-linked immunosorbent assay. Samples of blood were cultured (BACTEC) and incubated at 37°C for 5 days and then on Brucella agar. In addition, clinical and laboratory characteristics of brucellosis were done. The patients who had brucellosis (Coombs test ≥1/80 and 2ME ≥1/40) were selected. Data were analyzed using SPSS statistical package. Results: About 62.5% of patients were female with mean age of 37.5% years. 54.8% of cases were living in urban and 45.2% in rural areas. The most contagious seasons were spring. The most common transmission way was consuming of contaminated dairy products (59.3%); however, some of the patients had a history of animal contacts. The most common symptoms were fever (65.2%), arthralgia (68.1%), sweating (32%), malaise and fatigue (37.2%). The most common clinical signs were fever and peripheral arthritis. High leukocytosis and elevated erythrocyte sedimentation rate (>20 mm/h) were reported in all of the cases. Elevated C-reactive protein was detected in 72.1% of patients. Conclusion: Brucellosis should be considered in the differential diagnosis of patients with prolonged fever, spondylitis, or peripheral arthritis in endemic areas.