Abstract: Objective To investigate the effect of storage time on the accuracy of detection of human leukocyte antigen-B27(HLA-B27). Methods HLA-B27 FITC/CD3 PE double color fluorescent antibody staining was used, and flow cytometry was used to detect samples. The influence of storage time after staining and blood sample storage time on the detection results was compared. Results The average fluorescence intensity values detected by fluorescence 1 detection channel(FL1)at 0, 20, 40, 60, 80, 100, 120, 180 and 300 min after staining were(139.7±27.1),(136.6±26.2),(133.2±26.3),(130.9±25.3),(128.7±25.4),(127.3±25.1),(126.2±24.6),(122.4±24.0)and(116.6±22.1)respectively. With the extension of storage time, the average fluorescence intensity values of the samples decreased gradually, and the attenuation of the average fluorescence intensity was correlated with the storage time. The average fluorescence intensity values of blood samples at 0 min and 1, 2, 4, 8, 24 and 48 h were(139.7±27.1),(140.0±26.9),(140.1±27.3),(139.0±26.9),(140.3±26.0),(139.0±27.7)and(135.1±26.4)respectively. The result of blood samples stored for 48 h showed significant difference compared with that of fresh samples placed for 0 min(P<0.05). Conclusion The samples should be detected immediately after preparation, and blood samples should be detected within 24 hours after collection.