BACKGROUND Quantitation of peripheral blood (PB) CD34(+) cells is now an established method for timing PBPC harvesting. Recent refinements to the dual-platform ISHAGE gating strategy for CD34(+) cells has seen the introduction of microbeads to enable absolute counting of cells on a single instrument platform. This eliminates the need for total WBCC performed on an automated hematology analyzer and potentially increases the analytical precision of the methodology. At the same time, alternative methods for CD34(+) cell enumeration have started to emerge, notably microvolume fluorimetry, which forms the basis of the fully-automated STELLer CD34 method using the Imagn 2000. METHODS We performed a three-way evaluation of these methods. Sixty-eight samples of PB from 42 patients undergoing PBPC mobilization were analyzed by all three methods and correlations between all three calculated. The two-platform ISHAGE method was used as the reference method. RESULTS Precision and linearity of the single-platform and STELLer CD34 assays were excellent. Correlation with the dual-platform reference method was also excellent (single-platform method slope = 1.03, intercept = -0.03 and R(2) = 0.9325, STELLer CD34 assay slope = 0.827, intercept = 4.27, R(2) =0.8215). Bias, determined by Bland-Altman analysis, was 1.16 and -1.62 for single platform and STELLer CD34 assay respectively. CONCLUSION The three methods of CD34(+) cell enumeration gave equivalent results. The single-platform methodology negated the need for a separate white cell analyzer, while the STELLer CD34 methodology was technically the simplest.

中文翻译：

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外周血中 CD34+ 细胞计数的三种方法的比较：双平台 ISHAGE 协议与单平台，与微量荧光法

背景 外周血 (PB) CD34(+) 细胞的定量现在是一种确定 PBPC 采集时间的方法。最近对 CD34(+) 细胞的双平台 ISHAGE 门控策略进行了改进，引入了微珠，以便在单个仪器平台上对细胞进行绝对计数。这消除了在自动血液分析仪上执行总 WBCC 的需要，并有可能提高该方法的分析精度。与此同时，CD34(+) 细胞计数的替代方法已经开始出现，特别是微量荧光法，它构成了使用 Imagn 2000 的全自动 STELLer CD34 方法的基础。方法。来自 42 名接受 PBPC 动员的患者的 68 个 PB 样本通过所有三种方法进行分析，并计算所有三种方法之间的相关性。两平台ISHAGE方法被用作参考方法。结果 单平台和 STELLer CD34 检测的精密度和线性非常好。与双平台参考方法的相关性也很好（单平台方法斜率 = 1.03，截距 = -0.03 和 R(2) = 0.9325，STELLer CD34 测定斜率 = 0.827，截距 = 4.27，R(2) =0.8215） . 通过 Bland-Altman 分析确定的偏差对于单一平台和 STELLer CD34 测定分别为 1.16 和 -1.62。结论 CD34(+) 细胞计数的三种方法给出了相同的结果。单一平台方法不需要单独的白细胞分析仪，