标本前处理因素对流式细胞术检测淋巴细胞亚群的影响  

Effect of pre-analytical factors on detection of lymphocyte subset by flow cytometry

作　　者：费奇力[1] 王维维[1] 袁向亮[1] 张良[2] 

机构地区：[1]上海交通大学医学院附属新华医院检验科,上海200092 [2]上海交通大学医学院附属新华医院泌尿外科,上海200092

出　　处：《诊断学理论与实践》2014年第3期325-328,共4页Journal of Diagnostics Concepts & Practice

基　　金：国家自然基金青年项目(81301788)

摘　　要：目的：探讨抗凝剂、保存时间和保存温度等分析前因素对流式细胞术检测外周血淋巴细胞亚群的影响。方法：随机采集健康人全血10份,分别用乙二胺四乙酸二钾（EDTA-K2）和肝素钠2种抗凝剂抗凝,4℃及室温（18℃-22℃）保存,在不同时间点（〈4 h、24 h及48 h）分别使用荧光抗体标记,用流式细胞术检测CD3＋T淋巴细胞、CD3＋CD4＋辅助性T细胞、CD3＋CD8＋杀伤性T细胞、CD19＋B淋巴细胞及CD16＋CD56＋NK细胞占淋巴细胞的百分比,并采用荧光微球进行各淋巴细胞亚群的绝对计数。结果：与2种抗凝剂各自的室温条件下4 h内检测结果相比较,在肝素抗凝剂组中4℃保存至48 h,CD3＋T细胞、CD4＋T细胞、CD8＋T细胞、NK细胞的绝对计数显著降低（P〈0.05）;肝素抗凝血与EDTA-K2抗凝血间在4℃及室温保存至24 h及48 h的淋巴细胞亚群百分比差异均无统计学意义（P〉0.05）。结论：在检测总淋巴细胞亚群比例及绝对计数时,肝素钠抗凝的全血标本应保存在室温下,并在48 h内完成检测;EDTA-K2抗凝全血标本则可在4℃及室温下保存,在48 h内完成检测。Objective： To study the influence of anticoagulant, storage duration and storage temperature on detection of lymphocyte subset by flow cytometry. Methods： Ten blood samples were anticoagulated with EDTA-K2 and sodium heparin, respectively. Absolute counting detected by fluorescence labelling and percentages of lymphocyte subsets （CD3＋ T cells, CD4＋ T cells, CD8＋ T cells, CD19＋ B cells and NK （CD16＋CD56＋）cells determined by flow cytometry were analyzed at 〈4 h, 24 h, 48 h after storage under 4 ℃ or room temperature （18 ℃-22 ℃）. Results： Two kinds of anticoagulated samples in different conditions were compared with results of lymphocyte subsets detected within 4 h under room temperature. Absolute counting of CD3＋ T cells, CD4＋ T cells, CD8＋ T cells, NK cells in sodium heparin anticoagulated samples declined significantly after storage for 48 h under 4℃ （P〈0.05）. Percentages of lymphocyte subsets in different samples （different anticoagulants, different storage duration, different storage temperature） were similar. Conclusions： For absolute counting of lymphocyte subsets, EDTA-K2 anticoagulated samples could be stored either under 4 ℃ or room temperature and should be tested within 48 h. Sodium hepafin anticoagulated samples should be detected within 48 h when stored under room temperature.

关 键 词：流式细胞术 淋巴细胞亚群 分析前因素 抗凝剂 

分 类 号：R446.1[医药卫生—诊断学]