摘要
目的评价外周血B细胞上糖肌醇磷脂(glycosylphosphatidylinositol,GPI)连接蛋白之一CD59的表达能否作为诊断阵发性睡眠性血红蛋白尿症(paroxysmalnocturnalhemoglobinuria,PNH)的指标。方法采用流式细胞术双色免疫荧光分析法,结合B细胞特异性抗体CD20,分析92例外周血标本B细胞和中性粒细胞上CD59的表达,并通过临床指标评价方法,分析该指标临床应用的可行性。结果健康人B细胞上CD59均为完全阳性表达,表达率的参考范围为(96.3±1.2)%;PNH患者B细胞上CD59的表达率降低,均出现阴性或部分阳性病态细胞,病态细胞表达率为10.5%~92.3%;92.9%(26/28)非PNH贫血患者B细胞上CD59的表达均在正常参考范围内。该方法精密度的变异系数(coefficientvariation,CV)值小于4.4%,标本染色后无固定剂情况下可稳定保存24h,与常规检测CD59的方法(即中性粒细胞上CD59检测法)比较,PNH阳性和阴性检出符合率均高达100%。结论采用流式细胞术检测B细胞上CD59的表达具有简单、准确、重复性好的特点,可以成为诊断PNH的新指标。
To estimate whether the expression of glycosylphosphatidylinositol (GPI)-anchored protein in B lymphocytes could be an indicator for detecting paroxysmal nocturnal hemoglobinuria (PNH).Methods Flow cytometry and two -color McAbs were used to detect CD59expression in B lymphocytes and granulocytes of92clinical samples.The feasibility of the indicators for diagnosis of PNH was assessed.Results B lymphocytes from healthy individ uals mainly showed a single population of cells strongly positive for CD59,as the percentage of CD59expression was(96.3±1.2)%.In PNH patients,B lymphocytes showed decreased expression of CD59,as well as negative cells and partly-positive cells were observed,as the percentage of CD59expression was10.5%~92.3%.In26of28non-PNH anemia patients,CD59expression in B lymphocyte was normal,and in the other2patients,CD59expression were decreased(<95%),but no negative cell peak was observed.The precision CV of CD59in B lymphocyte assay is less than4.4%,stained samples could be kept stable for24h,the CD59in B lymphocyte assay was100%in accordance with routine assay(CD59in granulocyte assay).The sensitivity of the indicators(CD59in B lymphocytes and granulocytes)was100%,the specificity was97.4%,misdiagnosis rate was2.6%,the rate of failing to diagnosis was0%,the rate of positive forecast was87.5%,the rate of negative forecast was100%,the accuracy was97.8%.Conclusions The test of CD59expression in B lymphocytes by flow cytometry was simple,accurate and reproduceable.It could be a good marker for diagnosis of PNH because of high sensitivity and specificity.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2003年第5期599-602,共4页
Acta Academiae Medicinae Sinicae