流式细胞仪法用于血小板相关抗体检测和血小板交叉试验

目的 应用流式细胞仪(FCM)进行血小板相关抗体检测和血小板交叉试验。方法 利用血小板抗体阳性、阴性血清探寻FCM法实验条件,血小板经血清致敏,洗涤后,加入荧光标记鼠抗人IgG反应,由FCM获取和分析。52人次正常人血清经FCM法检测后,统计界定正常值范围。1份阳性血清经9个稀释度倍比稀释,分别用FCM法和固相ELISA抗体筛选法检测。31名临床血小板输注无效患者分别用FCM法和抗原捕获酶免分析法(MACE)进行102人次血小板交叉配合试验。结果 FCM法检测正常值为荧光强度比值R<1.156,9个稀释度的阳性血清检测表明,FCM法最高可检稀释度为86.67,固相ELISA法最高可检稀释度为56.34(抗-HPA)和67.25(抗-HLA)。102人次血小板交叉结果显示,FCM法和MACE法无显著性差异(P>0.05)。结论 FCM法可用于血小板相关抗体检测和血小板交叉配血,是一种快速、灵敏的新方法。

Detection of platelet-associated antibody and platelet crossmatching by flow cytometry

Objective Use of flow cytometry(FCM) in platelet - associated antibody detection and platelet cross-matching. Methods The condition of FCM assay for platelet-associated antibody was explored using a positive and a negative sera,and the cutoff value was calculated according to FCM assay of 52 normal sera and platelets. Nine serial delusions of 1 positive serum were tested by FCM assay and solid phase ELISA platelet antibody screening assay, and 102 platelet crossmatches were performed for 31 patients with platelet transfusions refractoriness by FCM assay and modified antigen-capture ELISA( MACE). Results The cutoff value of FCM assay is 1.156 for fluorescence ratio(R), and the highest dilution at which the antibody can be detected was 86.67 by FCM assay, 56.34 and 67.25 by solid phase ELISA for HPA antibody and HLA antibody. There was no difference in platelet crossmatch between FCM assay and the MACE assay(P> 0.05).Conclusion FCM assay can be used as a rapid and sensitive method for detecting platelet-associated antibody and platelet crossmatching.