Objective To improve clinicians' understanding on Prevotella bloodstream infection (BSI), reduce the rate of misdiagnosis and missed diagnosis, and broaden the ideas of diagnosis and treatment. Methods Clinical data of patients with Prevotella BSI at a hospital affiliated to a medical school of Nanjing University from May 2013 to May 2023 were collected. Risk factors, sources of infection, strains of infection, clinical manifestations, laboratory test results, treatment, and outcomes of patients with Prevotella BSI were retrospectively analyzed. Results A total of 23 patients diagnosed with Prevotella BSI were included in analysis, 15 (65.2%) were males and 8 (34.8%) were females. Most patients had related predisposing factors before BSI, such as surgical procedures (n=11, 47.8%), malignant tumors (n=10, 43.5%), diabetes (n=9, 39.1%), and indwelling urinary catheter (n=10, 43.5%), etc. There were 9 types of infected bacteria, mainly Prevotella buccalis (n=6, 26.1%), Prevotella bivia (n=5, 21.7%) and Prevotella intermedia (n=4, 17.4%). The main sources of infection were hepatobiliary system (n=6, 26.1%), abdominal and thoracic cavities (n=4, 17.4%), as well as urogenital tract (n=4, 17.4%). All patients showed symptoms of chills and fever, with significantly elevated blood inflammation indicators. Four cases (17.4%) developed septic shock, and 18 cases (78.3%) had a good prognosis after appropriate anti-infection treatment. Conclusion When atypical BSI caused by Prevotella is suspected, predisposing factors should be removed as soon as possible, blood should be actively collected and performed culture, rational use of antimicrobial agents based on antimicrobial susceptibility testing is beneficial for rapid control of infection and improvement of prognosis.