流式细胞术检测HLA-B27最佳抗体剂量与作用时间研究

The optimal dose and reaction time of HLA-B27 antibody in flow cytometry

目的探讨流式细胞术检测人类白细胞抗原B27(HLA-B27)时所用的最佳抗体剂量与作用时间。方法取52例强直性脊柱炎患者全血,根据HLA-B27抗体剂量不同分为5、10μL两组,分别避光静置5、10、15min,溶血后上机检测HLA-B27水平。结果 5μL抗体组不同反应时间HLA-B27阳性率分别为(84.16±1.21)%、(94.81±1.33)%、(94.1±1.26)%;10μL抗体组不同反应时间HLA-B27阳性率分别为(85.4±1.27)%、(96.76±1.31)%、(95.36±1.45)%;10μL抗体组HLA-B27检测结果高于5μL抗体组,差异有统计学意义(F=90.08,P<0.05);抗体反应10、15min HLA-B27检测结果高于抗体反应5min检测结果,差异有统计学意义(F=60.25,P<0.05);抗体反应10min HLA-B27检测结果与抗体反应15min检测结果比较差异无统计学意义(F=1.08,P>0.05)。结论 HLA-B27抗体剂量10μL,反应10min为最佳实验方案;HLA-B27抗体剂量、反应时间无交互作用。

Objective To study the optimal dose and reaction time of human leukocyte antigen B27（HLA‐B27）antibody in flow cytometry .Methods Take 52 cases of whole blood in patients with ankyl‐osing spondylitis（AS） .According to HLA‐B27 antibody doses ,samples were divided into two groups：5 μL group and 10 μL group .HLA‐B27‐positive rate were tested after 5 ,10 ,15 min , respectively .Results The HLA‐B27 positive rate of 5 μL group at different reaction time were （84 .16 ± 1 .21）% ,（94 .81 ± 1 .33）% ,（94 .10 ± 1 .26）% ;the positive rate of 10 μL group at different reaction time were （85 .40 ± 1 .27）% ,（96 .76 ± 1 .31）% , （95 .36 ± 1 .45）% .The positive rate of HLA‐B27 in 10 μL group was higher than 10 μL group（F=90 .08 ,P〈0 .05） .The positive rate of HLA‐B27 after reacting for 10 and 15 min were higher than that after reacting for 5 min（F=60 .25 ,P〈0 .05） .There was not statistically significantly different between the reaction time of 10 min and 15 min（F=1 .08 ,P〉0 .05） .Conclusion The opti‐mal dose and reaction time of HLA‐B27 antibody in flow cytometry are 10μL and 10 min;There is not any interaction between anti‐body dose and the reaction time of HLA‐B27 antibody .