流式细胞术分析户尘螨致敏的嗜碱性粒细胞活化试验及其临床意义

目的建立流式细胞术(FCM)分析嗜碱性粒细胞活化试验(BAT)的方法并评估其在过敏性疾病诊断中的意义。方法以CD63/CD203c/CD45抗体组合,建立用FCM分析户尘螨致敏的嗜碱性粒细胞脱颗粒的方法。以皮肤点刺试验(SPT)作为金标准方法,将FCM检测活化嗜碱性粒细胞与荧光酶联免疫吸附试验(FELISA)测定特异性IgE(sIgE)结果作比较,分析其临床应用价值。结果以CD45和CD203c联合设门,可以得到纯的嗜碱性粒细胞;CD63是活化嗜碱性粒细胞的最佳标志;Spearman相关系数显示,sIgE的Unicap分级与活化嗜碱性粒细胞呈中度相关;FCM检测活化嗜碱性粒细胞与FELISA测定sIgE结果在户尘螨过敏性疾病诊断中的差异无统计学意义(P>0.05),但前者的敏感度、符合率、阳性预测值以及阳性似然比各项指标都优于后者。结论通过FCM分析CD63的表达来确定活化嗜碱性粒细胞是诊断速发型超敏反应的一种有效、安全的体外检测方法。

Flow cytometry analysis of basophil degranulation in allergy to dust mite

Objective To establish the method for analysing basophil activation test(BAT) by flow cytometry(FCM) and evaluate its clinical significance in the diagnosis of allergic disease. Methods The protocol for FCM analysis of basophil degranulation in allergy to dust mite by CD63/CD203c/CD45 combination was established.The clinical significance of activated basophil by FCM was evaluated by comparing with the results of sIgE by fluorescence enzyme-linked absorbent assay(FELISA),regarding the skin prick test as the gold standard. Results Pure basophils were got by CD45 and CD203c gating;CD63 was the best marker for activated basophil;Spearman's correlation coefficients indicated a moderate positive correlation between SIgE class categorized by Unicap class and activated basophil;There was no significant difference between activated basophil by FCM and sIgE by FELISA in the diagnosis of allergic reaction to dust mite,but the former was better than the latter in sensitivity,accordance rate,positive predictive value and positive likelihood ratio. Conclusions Quantification of activated basophil by CD63 expression with FCM is a valuable new and safe method in vitro for diagnosis of immediate type hypersensitization.