Beckman Coulter三色荧光抗体干粉试剂检测CD4+T淋巴细胞的应用评价

Application of Beckman Coulter Three-Color Fluorescent Antibody Dry Reagent in CD4+T Lymphocyte Test

目的通过对三色荧光抗体干粉试剂(CD3/4/8)(Beckman Coulter,简称BC干粉试剂)应用于两种流式细胞仪(BD FACSCalbur和Beckman Coulter FC500)获得CD4+T淋巴细胞(简称CD4)检测结果的比较,从而评估其现场应用价值。方法 2015-11/12期间在四川省收集的198份HIV阳性静脉全血,以BD FACSCalibur流式细胞仪配套BD三色荧光抗体试剂(CD3/4/8)(简称BD试剂)作为参考方法,对BC干粉试剂在两种仪器上获得CD4结果的准确性和重复性进行评价。结果 BC干粉试剂在FACSCalbur、FC500流式细胞仪上获得CD4计数与参考方法结果的Pearson相关系数分别为0.84、0.95;平均偏差分别为-7个/μl(LOA:320.7~297.0个/μl)、5个/μl(LOA:180.3^-171.0个/μl);将CD4计数结果按≥500、350~500、200~350、<200个/μl进行分层,BC干粉试剂在两种仪器上的检测结果与参考方法比较无统计学差异(χ~2=0.431,P>0.05)、(χ~2=0.527,P>0.05);BC干粉试剂在两种仪器上获得CD4结果与参考方法无统计学差异(t=1.044,P>0.05)、(t=0.769,P>0.05);BC干粉试剂在两种流式细胞仪上检测的结果的平均偏差为-7个/μl(LOA:313.0^-326.6个/μl)。1~53号样本用BC干粉试剂分别在FACSCalbur和FC500流式细胞仪上重复检测1次,相同机型复检结果与第一次检测结果比较无统计学差异(t=0.26,P>0.05)、(t=0.235,P>0.05)。结论 BC干粉试剂应用于FACSCalbur和FC500流式细胞仪获得CD4结果具有较好的相关性和一致性,检测重复性较好。

Objective To evaluate the application value of three-color fluorescent antibody dry reagent（ CD3/4/8）（ Beckman Coulter）（ BC dry reagent） by comparing CD4＋T lymphocyte（ CD4） test results obtained with the reagent on two flow cytometers（ BD FACSCalbur and Beckman Coulter FC500）. Methods Accuracy and repeatability of CD4 results obtained with BC powder reagent on the two instruments were evaluated on 198 HIV positive blood samples collected in Sichuan Province during November and December of 2015,using the result obtained with a BD FACSCalibur flow cytometer and BD three color fluorescent antibody reagents（ CD3/4/8）（ BD reagents） as reference. Results The Pearson correlation coefficient between CD4 counts obtained with the BC powder on FACSCalbur, FC500 flow cytometers and that obtained with the reference method were 0. 84 and 0. 95,respectively; the average deviation is-7 cells/L（ LOA： 320. 7 to-297. 0 cells/L） and 5 cells/L（ LOA： 180. 3 to-171. 0 cells/L）,respectively. The CD4 count results re stratified into ≥500,350 to 500,200 to 350 and 200 cells/μl layers,and no statistical difference was found between results obtained with BC dry powder reagent on the two instruments and those obtained with the reference method（ χ~2= 0. 431,P〈0. 05）,（ χ~2= 0. 527,P〈0. 05）. No statistical difference was found between CD4 counts obtained with BC dry powder reagent on the instruments and that obtained with the reference method（ t = 1. 044,P〈0. 05）,（ t = 0. 769,P〈0. 05）; the average deviation of results obtained with BC dry reagent on the two flow cytometers was-7 cells/L（ LOA： 313. 0 to-326. 6 cells/L）. Samples 1 to 53 were repeatedly tested with BC reagent on FACSCalbur and FC500 flow cytometers,and no statistical difference was found between test and retest results obtained with the same instrument model（ t =0. 26,P〈0. 05）,（ t = 0. 235,P〈0. 05）. Conclusion With BC dry powder reagent on FACSCalbur and FC500 flow cytometers,the CD4 results obtained are well correlated and consistent,and the test is well repeatable.