标本存放时间及温度对刺激法Th1/Th2胞内细胞因子检测的影响

目的探讨标本不同存放时间及温度对刺激法检测外周血Th1/Th2胞内细胞因子检验结果的影响。方法对20例乙二胺四乙酸(EDTA)抗凝外周血全血标本放置4℃冰箱及室温(20±5)℃保存,在不同的时间点取出标本,经过PMA+Ion刺激后采用流式细胞诊断技术检测Th1(CD4⁺/IFN-γ⁺)和Th2细胞(CD4⁺/IL-4⁺),计算Th1/Th2值,对不同温度和不同存放时间的检测结果进行比较。结果标本室温或4℃冰箱放置时,在72 h内检测结果相对稳定,检测结果变化差异无统计学意义(P>0.05)。标本存放3 d后,无论是室温放置,还是在冰箱存放都与抽血当天检测结果比较存在较大的差异(P<0.05)。结论放置时间是影响检验结果的主要因素,而存放温度无明显影响。

Effects of time and temperature of specimen storage on the results of cytokine Th1/Th2 in peripheral blood

Objective To investigate the effects of different storage time and temperature on the results of Th1/Th2 test in peripheral blood. Methods Twenty cases of ethylenediaminetetraacetic acid (EDTA) anticoagulant peripheral blood samples were stored in refrigerator at 4℃and room temperature (20±5)℃. The samples were taken out at different time. After stimulated by PMA + Ion, Th1 (CD4+/IFN-γ+) and Th2 cells (CD4+/IL-4+), the samples were detected by flow cytometry; and Th1/Th2 values were calculated and the detection results at different temperature and storage time were compared. Results When the samples were placed at room temperature or 4℃refrigerator, the results were relatively stable within 72 hours, and there was no significant difference in the results (P>0.05). After 3 days, there were significant differences between the test results of samples stored at room temperature or in refrigerator and the first day of blood sampling. Conclusion The storage time is the main influence factor on the test results, while the storage temperature has no obvious effect.