Clinical Department of Transfusion Medicine and Transplantation Biology, University Hospital Centre Zagreb, Zagreb, Croatia.
Int J Lab Hematol. 2021 Feb;43(1):61-67. doi: 10.1111/ijlh.13315. Epub 2020 Sep 15.
CD34+ hematopoietic stem cell (HSC) enumeration by cell flow cytometry is routinely used in clinical laboratories for monitoring of HSC mobilization into peripheral blood and assessment of the quality of HSC products. The modified ISHAGE protocol is the most often used procedure for determination of CD34+ cells using flow cytometry. The aim of this study was to evaluate BD Enumeration stem cell kit on flow cytometer BD facscanto II, using facscanto clinical and facsdiva softwares.
Validation study included determination of within-run and between-run precision, trueness (bias), comparison of the test results analyzed on facscanto clinical and facsdiva softwares, assessment of linearity, specimen stability, and carryover.
For between-run precision, coefficients of variation (CVs) were all <10%, except for low control level on facsdiva software. CVs for within-run precision were <10%, except for high absolute count of CD34+ cells on facsdiva software. Comparison of data showed no statistically significant differences between facscanto clinical and facsdiva software (Spearman's rank correlation coefficients were .993 for % of CD34+ cells and 0.983 for absolute count of CD34+ cells). In linearity study, bias for all dilutions was < 20%, and carryover assessment cannot be considered significant on both softwares. There was a statistically significant difference (P = .044) in absolute count of CD34+ cells after 24 hours of storage, when using facscanto clinical software.
BD Stem Cell Enumeration Kit can be used in routine laboratory work on BD FACSCanto II instrument, whereas facscanto clinical and facsdiva software were used for acquisition and data analysis.
通过细胞流式细胞术对 CD34+造血干细胞（HSC）进行计数，通常用于临床实验室以监测 HSC 动员到外周血中，并评估 HSC 产品的质量。改良的 ISHAGE 方案是最常用于通过流式细胞术确定 CD34+细胞的程序。本研究旨在使用流式细胞仪 BD facscanto II 上的 BD 计数干细胞试剂盒评估 facscanto clinical 和 facsdiva 软件。
验证研究包括确定批内和批间精密度、准确性（偏差）、比较在 facscanto clinical 和 facsdiva 软件上分析的测试结果、评估线性、标本稳定性和携带污染。
对于批间精密度，除了 facsdiva 软件的低对照水平外，变异系数（CV）均<10%。批内精密度的 CV<10%，除了 facsdiva 软件上 CD34+细胞的绝对计数较高外。数据比较显示，facscanto clinical 和 facsdiva 软件之间无统计学差异（Spearman 等级相关系数分别为 CD34+细胞的%为.993 和 CD34+细胞的绝对计数为 0.983）。在线性研究中，所有稀释度的偏差均<20%，在这两个软件上都不能认为携带污染有显著影响。使用 facscanto clinical 软件时，24 小时储存后 CD34+细胞的绝对计数存在统计学差异（P=0.044）。
BD Stem Cell Enumeration Kit 可在 BD FACSCanto II 仪器的常规实验室工作中使用，而 facscanto clinical 和 facsdiva 软件用于采集和数据分析。
