人淋巴细胞国家参考品的研制  

Establishment of the national reference materials of human lymphocytes

作　　者：胡泽斌[1] 高飞[1] 曲守方[1] 孙彬裕[1] 杨振[1] 孙晶 黄杰[1] HU Ze-bin;GAO Fei;QU Shou-fang;SUN Bin-yu;YANG Zhen;SUN Jing;HUANG Jie(National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国食品药品检定研究院体外诊断试剂检定所,北京100050

出　　处：《中国医药生物技术》2020年第5期475-481,共7页Chinese Medicinal Biotechnology

基　　金：国家科技重大专项(2018ZX10732401-003-009)。

摘　　要：目的制备人淋巴细胞国家参考品,用于人T/B/NK淋巴细胞亚群检测试剂的准确性评价。方法收集健康志愿者的新鲜外周血白细胞,采用人淋巴细胞分离液无菌分离得到外周血单个核细胞(PBMCs)。将PBMCs均匀重悬于细胞固定液,于2~8℃静置过夜。次日离心去除细胞固定液,加入适量冻干保护液重悬细胞。每支西林瓶按1 ml进行分装,分装后的细胞进行冻干,制备成人淋巴细胞国家参考品候选品。同时对候选品的分装均匀性、瓶间均匀性、基质效应和稳定性进行评价。结果成功研制出冻干型人淋巴细胞国家参考品,其均匀性和稳定性良好且无基质效应。结论人淋巴细胞国家参考品候选品各项指标均符合要求,可用于人T/B/NK淋巴细胞亚群检测试剂(流式细胞法)的准确性评价。Objective To prepare the national reference materials of human lymphocytes for evaluation of the accuracy of human T/B/NK lymphocyte subsets detection reagents.Methods Peripheral blood mononuclear cells(PBMCs)were isolated from fresh peripheral blood leukocytes of healthy volunteers.The PBMCs were re-suspended in the cell fixed solution,and placed overnight at 2-8℃.On the next day,the fixed solution was removed by centrifugation,then the cells were re-suspended in lyophilization protective solution.Each vial was divided into 1 ml-vial,and the cells were lyophilized to prepare the national reference candidates.At the same time,the homogeneity between various vials and matrix effects along with stability of the candidates were evaluated.Results The national reference materials of lyophilized human lymphocytes with good homogeneity and stability were successfully developed.No matrix effects were observed.Conclusion All indexes of the national reference sample of human lymphocyte meet the requirements.The lymphocytes can be used for the accuracy evaluation of human T/B/NK lymphocyte subsets detection reagent(flow cytometry).

关 键 词：淋巴细胞 国家参考品 流式细胞法 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]