The invention discloses a method for indirect immunofluorescence assay (IFA) detection of eperythrozoonosis by use of a monoclonal antibody. The method includes establishment of a standard positive control and a standard negative control, antigen coating, fluorescence second antibody dilution, IFA determination and other steps. An antibody used in the method is a monoclonal antibody against porcine eperythrozoon, and compared with general antibodies used in IHA (indirect hemagglutination assay), ELISA (enzyme-linked immuno sorbent assay) and other detection methods, the antibody used in the method has the advantages of high specificity, high sensitivity, high accuracy and reliability, can fast, simply, reliably and accurately detect eperythrozoon in a sample, and can also be used for epidemiological investigation and curative effect detection of the eperythrozoonosis.