Objective To study the method of laboratory Brucella detection and evaluate its clinical efficacy. Methods The clinical case of Brucella infection in sheep diagnosed by medical staff in our hospital in March 2019 was taken as the research object. The advantages and disadvantages of serum agglutination test, routine blood culture and bacterial identification, and mass spectrometry identification methods in diagnosing Brucella were analyzed and discussed, providing some reference for the detection of Brucella in grassroots hospital laboratories. Results The medical staff's serum brucellosis tiger red plate agglutination antigen test was positive. The blood culture BD PhoenixTM whole bacterial identification/drug sensitivity system instrument in the United States indicates that the aerobic bottle reports positive, but cannot identify the species of Brucella. The mass spectrometer identified it as Brucella lamblia. Conclusion The above methods have their own advantages and disadvantages in application, grassroots hospital laboratories should choose suitable methods for detecting Brucella based on actual conditions.