To provide a theoretical basis for the early diagnosis and treatment of brucellosis in Honghe Prefecture, Yunnan Province, this study analyzed the clinical characteristics of confirmed cases of brucellosis in Yunnan region, and compared these cases with the reported cases in the literature. The clinical data of 36 patients with brucella infection confirmed by positive blood culture admitted from Jan. 2018 to Dec. 2022 were retrospectively analyzed. Based on the clinical characteristics and diagnosis, they were divided into classic fever of unknown origin (FOU) group and non classic fever of unknown origin (non-FOU) group, and the epidemiological and clinical characteristics of the two groups were analyzed. The results demonstrated that, 91.67% patients lived mainly in rural areas; the acute stage is mainly, accounting for 88.89%, fever is one of the most common clinical symptoms, accounting for 94.44%, and 80.56% were complicated with liver injury. Comparing the epidemiological characteristics, clinical features, laboratory test results, and clinical outcomes of the two groups of patients, the non-FUO group had a significantly higher misdiagnosis rate than the FUO group (94.74% vs. 64.71%, P＝0.023); the proportion of fever accompanied by chills in the FUO group was significantly higher than that in the non-FUO group (77.78% vs. 22.22%, P＝0.000). There was no statistical difference between the two groups in the remaining data (P>0.05). It was concluded that, brucellosis have a high misdiagnosis rate, and patients with non-FUO are more likely to be misdiagnosed as other diseases. Incidents of Brucella infection in the Honghe of Yunnan are sporadic throughout the year, and doctors need to attach great importance to inquiring about epidemiological history and relevant laboratory tests to reduce the missed diagnosis and misdiagnosis rate of the disease.