Center for Biologics Evaluation and Research, Food & Drug Administration, 8800 Rockville Pike, Bethesda, MD 20892, USA.
Cytometry B Clin Cytom. 2011 Sep;80(5):300-8. doi: 10.1002/cyto.b.20591. Epub 2011 Apr 6.
ZAP-70 expression is a stage independent prognostic marker in CLL. However, interlaboratory variation is large, and there is neither a consensus nor a regulatory approved methodology.
Two anti-ZAP70 clones (1E7.2 and SBZAP) were compared in 45 untreated CLL patients. Nine different methods for ZAP-70 expression analysis were evaluated: M1, isotype control to determine negative; M2, internal residual T-cell to determine positive; M3, normal donor (ND) T-cell to determine positive; M4, internal T-cell/clone ratio; M5, ND residual T-cell/clone ratio; M6, clone/normal remaining B-cell ratio; M7, clone/ND B- cell ratio; M8, CLL-Z score; M9, modified CLL-Z score. A scoring system was designed integrating both 1E7.2 and SBZAP clones to assign ZAP-70 expression.
The correlation coefficients for the four selected highest statistically significant methods were as follows (M1 = 0.71, M3 = 0.72, M7 = 0.67, and M9 = 0.64). These four methods were used to generate a combined score. The two reagents showed agreement using the designed scoring system for 37/45 samples (82%), and 8/45 (18%) showed equivocal result with one of the two clones. Seven of the eight equivocal samples were resolved using the scoring system.
Four of the nine methods of analysis were compared for each reagent. The use of two independent ZAP-70 reagents increases analytical certitude and the scoring method aids in the resolution of equivocal results. The combined use of two reagents, four methods of analysis, and a scoring method allowed for assignment of ZAP-70 expression in 44/45 samples (98%) tested and improved performance of this important prognostic assay.
ZAP-70 表达是 CLL 中独立于分期的预后标志物。然而，实验室间差异较大，目前既没有共识也没有监管批准的方法。
在 45 例未经治疗的 CLL 患者中比较了两种抗 ZAP70 克隆（1E7.2 和 SBZAP）。评估了 9 种不同的 ZAP-70 表达分析方法：M1，同种型对照确定阴性；M2，内源性残留 T 细胞确定阳性；M3，正常供体（ND）T 细胞确定阳性；M4，内源性 T 细胞/克隆比；M5，ND 残留 T 细胞/克隆比；M6，克隆/正常剩余 B 细胞比；M7，克隆/ND B 细胞比；M8，CLL-Z 评分；M9，改良 CLL-Z 评分。设计了一种评分系统，整合了 1E7.2 和 SBZAP 两种克隆，用于分配 ZAP-70 表达。
四种选择的统计学上最显著方法的相关系数如下（M1 = 0.71，M3 = 0.72，M7 = 0.67，M9 = 0.64）。使用这四种方法生成了一个综合评分。这两种试剂使用设计的评分系统对 37/45 个样本（82%）的结果一致，对其中一种试剂有 8/45（18%）的结果不确定。使用评分系统解决了 8 个不确定样本中的 7 个。
对每种试剂比较了 9 种分析方法中的 4 种。使用两种独立的 ZAP-70 试剂增加了分析的确定性，评分方法有助于解决不确定结果。两种试剂、四种分析方法和一种评分方法的联合使用，能够对 44/45（98%）的检测样本进行 ZAP-70 表达的分配，从而提高了这个重要预后检测的性能。
