应用流式细胞术检测血小板聚集及活化功能临床观察

Evaluation of platelet function by flow cytometry

目的:探讨应用流式细胞术全血法检测血小板功能、影响因素及临床意义。方法:应用血小板特异性荧光抗体CD61-FITC标记血小板,并以0.82μm标准微球进行定位对照和设置机器检测条件,调节机器阈值,设门计数血小板微颗粒(PMP)占CD61阳性颗粒的百分比。以ADP及胶原诱导血小板活化,计数活化以后血小板释放的PMP,以未加血小板激活剂的标本作为零聚集对照,激活剂活化的标本以单个血小板数量的减少反应血小板聚集率的多少,并进行方法学的评价。结果:该方法能够有效的检测PMP及低浓度诱导剂诱导下的血小板聚集,检测的血小板聚集功能敏感度明显高于比浊法。应用枸橼酸钠和CTAD抗凝样本,检测结果显示CTAD抗凝的全血静息状态下血小板释放PMP的量显著低于枸橼酸钠抗凝全血释放PMP量。结论:流式细胞术全血法检测血小板功能方法准确、敏感,快速、简便适合于临床常规检测。

Objective：To determine the platelet function directly in whole blood by flow eytometry （FCM） and and to discuss its influence factors and clinical significance.Methods：Whole blood was prepared as samples and then were stained with a fluorochmme-conjugated monoclonal antibody directed towards platelet surface glycoproteins.O.82 p,m latex beads were used to calibrate of forward scatter threshold parameter and setup optimization of procedure of flow cytometry and as standard for counting PMP.PMP generated in vitro using ADP or collagen were detected as positive control,the single platelet as negative control for platelet aggregates and systemically evaluated the method.Results ：It was feasible using flow cytometry to evaluate the extent of platelet activation and the presence of PMP and platelet aggregates.Samples collected into two different anticoagulant acid-citrate-dextrose （ACD） and CTAD.The resting PMP of them were remarkahly different.Conclusion：The method is simple, sensitive and accurate with relatively less influence factor which can be as routine test.