摘要
目的探讨保存温度、保存时间对血清样本中白介素6(IL-6)、白介素8(IL-8)、白介素10(IL-10)、肿瘤坏死因子α(TNF-α)测定的影响。方法随机选取北京协和医院就诊的120例患者,根据保存条件分为5组:室温原管组40例,室温原管保存;室温分装组20例,血清分至EP管室温保存;4℃原管组20例,4℃保存不分装;4℃分装组20例,血清分至EP管4℃保存;-20℃组20例,血清分至EP管-20℃保存。室温原管组中的20例标本和室温分装组、4℃原管组、4℃分装组分别测定即刻、6 h、24 h的IL-6、IL-8、IL-10、TNF-α;室温原管组中的另20例标本用于分析IL-8的稳定性,分别测定即刻、2 h、3 h、4 h、5 h的IL-8;-20℃组测定即刻及-20℃冷冻6个月后的IL-6、IL-8、IL-10、TNF-α。结果室温原管组中IL-6、IL-10、TNF-α在6 h、24 h结果与即刻相比,差异均无统计学意义,IL-8在6 h、24 h结果与即刻相比,差异均有统计学意义(P<0.05),比对通过率分别为5%、0%,假阳性率分别为60%、70%。室温放置2 h、3 h、4 h、5 h IL-8的比对通过率分别为80%、35%、15%、10%,P值分别为0.820、0.461、0.076、0.003,仅2 h的结果通过稳定性比对。虽然其他4组各时间点细胞因子的测定结果与即刻相比差异均无统计学意义,但室温分装组IL-8在6 h、24 h的比对通过率分别为65%、50%;4℃原管组的IL-8在24 h的比对通过率为55%,均未通过比对。只有4℃分装组和-20℃组细胞因子各时间点的测定结果与即刻相比,差异均无统计学意义,且各项目比对通过率均≥80%,结果稳定可靠。结论采用血清样本联合检测IL-6、IL-8、IL-10、TNF-α时,分析前时间控制在2 h内,可以采取室温原管放置;2~24 h内推荐4℃分装;-20℃时6个月结果稳定。各实验室若不能满足2 h内及时上机检测,为避免IL-8的假阳性,建议分装冻存。
Objective To explore the influences of temperature and time in storage on the determination for interleukin-6(IL-6),interleukin-8(IL-8),interleukin-10(IL-10)and tumor necrosis factor-α(TNF-α)in serum samples.Methods A total of 120 serum samples from patients were randomly selected from Peking Union Medical College Hospital and divided into 5 groups according to storage conditions as follows:room temperature original tube group:40 serum samples were stored in original tubes at room temperature;room temperature subpackage group:20 samples were divided into EP tubes and stored at room temperature;4℃C original tube group:20 samples in original tubes were stored at 4℃C without subpacage;4 C subpackage group:20 samples were divided into EP tubes and stored at 4℃;-20℃group:20 samples were divided into EP tubes and stored at-20℃.The levels of IL-6,IL-8,IL-10 and TNF-αin 20 samples from the each group:room temperature original tubes,room temperature subpackage tubes,4 C original tubes and 4℃subpackage tubes were measured immediately,6 hours and 24 hours,respectively.The levels of IL-8 of the serum samples in the other 20 tubes stored in room temperature were measured immediately,2,3,4 and 5 hours respectively to analyze the stability of test results.The levels of IL-6,IL-8,IL-10 and TNF-αin the samples stored at-20 C group were tested immediately and after freezing for 6 months respectively.Results For the samples of room temperature original tube group,the results of IL-6,IL-10 and TNF-αmeasured at 6 h and 24 h do not showed statistically significant different compared with those measured immediately,but the results of IL-8 measured at 6 and 24 hours were both significantly different from the results measured immediately(P<0.05).The comparisonable pass rate were 5%and 0%,and the false positive rate were 60%and 70%,respectively.The comparisonable pass rates of the results of IL-8 measured after 2,3,4 and 5 hours at room temperature were 80%,35%,15%and 10%,respectively with the P values of 0.820,0.461,0.076 and 0.003 respectively.Only the results of IL-8 measured in 2 hours passed the stability comparison.Although there were no statistically significant difference for the measurement results of the four kinds of cytokins at each time point compared with those measured immediately,but the comparisonable pass rates of IL-8 results in the room temperature subpackage group at 6 h and 24 h were only 65%and 50%respectively,and the comparisonable pass rate of IL-8 results in original tube group at 4 C within 24 hours was 55%,both of which did not pass the comparison.Only the measurement results of the four cytokins at each time point in 4 C subpackaging group and-20 C group showed no statistically significant difference compared to the immediately measured results,and the pass rate of comparison were≥80%for each cytokin,indicating stable results.Conclusion The simultaneous combined determination for cytokines IL-6,IL-8,IL-10 and TNF-αshould control the pre-analysis time within 2 hours,but the sample tubes could be placed at room temperature.It is recommended to pack at 4 C within 2 h to 24 h.The test results of the sample stored at-20℃may remain stable for 6 months.If the laboratory can not attain to measure the samples timely on analytical instrument within 2 hours,it is recommended to repackage for cryopreserving the serum samples in order to avoid false positive results of IL-8.
作者
毛镭篥
张丽
李薇
黄媛
王斐
吴卫
MAO Leili;ZHANG Li;LI Wei;HUANG Yuan;WANG Fei;WU Wei(Department of Clinical Laboratory,Chinese Academy of Medical Science&Peking Union Medical College Hospital,Beijing 100730,China)
出处
《临床检验杂志》
CAS
2023年第6期411-416,共6页
Chinese Journal of Clinical Laboratory Science
关键词
细胞因子
血清样本
温度
时间
保存条件
cytokines
serum sample
temperature
time
preservation condition
作者简介
毛镭篥,1989年生,女,主管技师,硕士,主要从事临床检验与细胞学工作;通信作者:吴卫,副主任技师,E-mail:ww_pumch@sina.com。
