Van Bockstaele Femke, Janssens Ann, Piette Anne, Callewaert Filip, Pede Valerie, Offner Fritz, Verhasselt Bruno, Philippé Jan
Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
Cytometry B Clin Cytom. 2006 Jul 15;70(4):302-8. doi: 10.1002/cyto.b.20122.
ZAP-70 has been proposed as a surrogate marker for immunoglobulin heavy-chain variable region (IgV(H)) mutation status, which is known as a prognostic marker in B-cell chronic lymphocytic leukemia (CLL). The flow cytometric analysis of ZAP-70 suffers from difficulties in standardization and interpretation. We applied the Kolmogorov-Smirnov (KS) statistical test to make analysis more straightforward.
We examined ZAP-70 expression by flow cytometry in 53 patients with CLL. Analysis was performed as initially described by Crespo et al. (New England J Med 2003; 348:1764-1775) and alternatively by application of the KS statistical test comparing T cells with B cells. Receiver-operating-characteristics (ROC)-curve analyses were performed to determine the optimal cut-off values for ZAP-70 measured by the two approaches. ZAP-70 protein expression was compared with ZAP-70 mRNA expression measured by a quantitative PCR (qPCR) and with the IgV(H) mutation status.
Both flow cytometric analyses correlated well with the molecular technique and proved to be of equal value in predicting the IgV(H) mutation status. Applying the KS test is reproducible, simple, straightforward, and overcomes a number of difficulties encountered in the Crespo-method.
The KS statistical test is an essential part of the software delivered with modern routine analytical flow cytometers and is well suited for analysis of ZAP-70 expression in CLL.
ZAP - 70已被提议作为免疫球蛋白重链可变区（IgV(H)）突变状态的替代标志物，而IgV(H)突变状态是B细胞慢性淋巴细胞白血病（CLL）中的一种预后标志物。ZAP - 70的流式细胞术分析在标准化和解读方面存在困难。我们应用柯尔莫哥洛夫 - 斯米尔诺夫（KS）统计检验以使分析更直接。
我们通过流式细胞术检测了53例CLL患者的ZAP - 70表达。分析最初按照Crespo等人（《新英格兰医学杂志》2003年；348:1764 - 1775）所描述的方法进行，另外还通过应用KS统计检验将T细胞与B细胞进行比较。进行了受试者操作特征（ROC）曲线分析以确定通过两种方法测量的ZAP - 70的最佳临界值。将ZAP - 70蛋白表达与通过定量聚合酶链反应（qPCR）测量的ZAP - 70 mRNA表达以及IgV(H)突变状态进行比较。
两种流式细胞术分析均与分子技术密切相关，并且在预测IgV(H)突变状态方面被证明具有同等价值。应用KS检验具有可重复性、简单、直接，并且克服了Crespo方法中遇到的一些困难。
KS统计检验是现代常规分析流式细胞仪所配备软件的重要组成部分，非常适合分析CLL中ZAP - 70的表达。
