国产T淋巴细胞检测试剂和仪器应用评价

Accuracy of domestic T lymphocytes reagent and flow cytometry:an applicative evaluation

目的获得国产通用型CD4^+T淋巴细胞(CD4)检测试剂应用于开放型流式细胞仪检测结果的准确性。方法于2014年7月1-30日对艾滋病病毒(HIV)感染者样本全血样本206份进行平行检测,以BD 4色试剂配套BD FACSCalibur流式细胞仪检测结果为参考,评价同生3色(CD3/8/4、CD3/4/45)、4色试剂(CD3/8/45/4)在ACEA NovoCyte流式细胞仪获得T淋巴细胞亚群绝对数的准确性。结果206份全血样本分析结果如下:同生试剂各参数与参考方法相关系数均>0.96。同生3色(CD3/8/4)、(CD3/4/45)和4色(CD3/8/45/4)检测CD4绝对数的平均偏差分别为(2.49±50.5)、(–3.1±43.8)和(–6.2±58.2)个/μL,与参考方法相比均无统计学差异;CD3绝对数的平均偏差分别为(10.3±115.3)、(–1.7±108.0)和(14.3±125.4)个/μL,均无统计学差异;CD8绝对数平均偏差分别为(–4.6±78.6)和(–19.6±87.2)个/μL。结论同生试剂应用于ACEA NovoCyte流式细胞仪均具有良好的准确性,可用于常规T淋巴细胞检测。

Objective To evaluate the accuracy of a domestic reagent(Tongsheng reagent)for determination of absolute count of cluster of differentiation 4 positive(CD4^+)T lymphocytes with different multicolor flow cytometry.Methods Parallel tests for accuracy in absolute counts of T lymphocyte subgroups detected using domestic reagents and instrument were performed on whole blood samples of 206 human immunodeficiency virus(HIV)infected people during July 2014.The domestic reagents used in the detection were Tongsheng three color(CD3-FITC/CD8-PE/CD4-APC[CD3/8/4]/CD3-FITC/CD4-PE/CD45-PerCP[CD3/4/45])and Tongsheng four color(CD3-FITC/CD8-PE/CD45-PerCP/CD4-APC[CD3/8/45/4])and the domestic instrument was ACEA NovoCyte flow cytometry.The reference reagent and instrument were BD four color reagent kit and ral BD FACSCalibur flow cytometry.Results For all the blood samples,the correlation coefficients between the results from detections with domestic reagents and instrument and reference reagent and instrument were all greater than 0.96.Compared with the results using reference reagent and instrument,when using Tongsheng three color(CD3/8/4 or CD3/4/45)and Tongsheng four color(CD3/8/45/4)reagents and ACEA NovoCyte flow cytometry,the mean deviations of absolute counts(cell/μL)were 2.49±50.5,–3.1±43.8 and–6.2±58.2 for CD4;10.3±115.3,–1.7±108.0 and 14.3±125.4 for CD3;–4.6±78.6 and–19.6±87.2 for CD8,respectively.No significant differences were observed between the absolute counts of CD4,CD3,and CD8 T lymphocytes determined with domestic and reference reagents and instrument.Conclusion Domestic Tongsheng three color and four color reagents and ACEA NovoCyte can be used for accurate determinations of T lymphocytes absolute counts.