RhCE表型对RhD抗原量及抗-D效价测定的影响研究  

Study on the effect of RhCE phenotype on RhD antigen amount and anti-D titer determination

作　　者：赵俸涌[1] 王中英[1] 徐婷[1] 王晨[1] 郭忠慧[1] 李勤[1] 张嘉敏[1] 向东[1] 朱自严[1] ZHAO Fengyong;WANG Zongying;XU Ting;WANG Chen;GUO Zhonghui;LI Qin;ZHANG Jiamin;XIANG Dong;ZHU Ziyan(Shanghai Blood Center,Shanghai 200051,China)

机构地区：[1]上海市血液中心,上海200051

出　　处：《中国输血杂志》2020年第9期905-908,共4页Chinese Journal of Blood Transfusion

基　　金：上海市卫健委青年课题(20184Y0281);中国输血协会威高基金项目(CSBT-WG-2019-08)。

摘　　要：目的目前对Rh系统抗-D的定量或效价检测(相对定量)方法皆依赖于试剂红细胞。日常工作中,绝大多数实验室采用单人份红细胞作为效价测定的试剂红细胞,若RhCE表型对RhD抗原表达量影响显著,则不同RhCE表型必然会产生不同的抗-D效价测定结果。本研究旨在通过RhCE分型,RhD抗原定量试验及相应抗体进行效价测定,比较分析RhCE表型对RhD抗原量及抗-D效价测定的影响。方法采用血清学方法筛选获得CCee、Ccee、ccEE、CcEe表型的RhD阳性标本,并使用流式细胞术检测上述红细胞RhD抗原量,最后使用不同RhCE表型单人份标本红细胞对同一份抗-D进行效价测定。分析RhCE表型对RhD抗原量及抗-D效价测定的影响。结果在99例细胞红细胞标本进行Rh血型血清学分型后,对CCDee(n=17)、CcDee(n=15)、CcDEe(n=8)、ccDEE(n=8)分型细胞进行流式免疫荧光检测,发现CcDee与ccDEE及CcDEe间、CCDee与ccDEE之间RhD抗原的荧光量比较存在较大统计学意义差异(P<0.001),CcDee与CCDee比较存在统计学意义差异(P<0.01),其余各组间均不具统计学意义的差异;再对其中CCDee(n=15)、CcDee(n=14)、CcDEe(n=8)、ccDEE(n=8),使用同1份抗-D血浆进行效价测定分析,将效价结果与红细胞RhD抗原免疫荧光值(MFI)进行比较后,发现红细胞表面RhD抗原量与效价二者呈现正相关关系(r=0.907;P<0.001),对效价与RhD抗原MFI值进行统计分析发现:对同一份抗-D血浆,效价值为32、64、128各组内红细胞MFI具有统计学意义的差异(P<0.05);将效价与RhCE分型进行统计分析,发现CCEE与Ccee间、CcEe与Ccee间存在统计学意义的差异(P<0.05)。结论不同的RhCE表型会造成D抗原表达差异,D抗原量对效价测定有较大影响,提示在效价测定试验中,使用相同RhCE分型红细胞作为试剂细胞或对RhD抗原进行定量分析可提高效价试验的准确度。Objective Either the anti-D quantification or titration(relative quantification) methods relys on the RhD antigen on the reagent RBCs.The presence of RhCE antigen may affect the expression of RhD antigen, which in turn affects the results of titer determination. In daily work, most laboratories use a single donor′s RBC as a reagent cell for titter determination. If the RhCE phenotype has a significant effect on the expression of RhD antigen, it will inevitably affect the result of the titter determination. The purpose of this study was to clarify the relationship between the quantitation of D antigen on red blood cells and the typing of RhCE antigen and its effect of plasma anti-D titration.Methods Serological methods were used to screen RhCE typing and select RhD positive RBCs with CCee, Ccee, ccEE, CcEe phenotypes. Flow cytometry was adopted to detect the RhD antigen amount of above-mentioned RBCs, and anti-D titration of the same plasma sample was measured using RBCs with different RhCE phenotypes collected from different individuals. The effect of RhCE phenotypes on anti-D titration and the amount of antigen was evaluated.Results After Rh blood group serological classification of 99 cases of erythrocyte samples, 17 cases of CCDee, 15 of CcDee, 8 of CcDEe, and 8 of ccDEE were randomly selected and detected by flow cytometry, and significant differences were found in median fluorescence intensity(MFI) as CcDee in comparison to ccDEE and CcDEe, as well as CCDee in comparison to ccDEE(P<0.001), and statistical difference was found between CcDee and CCDee(P<0.01), while no other statistical differences were observed among other pairwise comparisons. 15 cases of CCDee, 14 of CcDee, 8 of CcDEe and 8 of ccDEE were randomly selected and analyzed by anti-D titration using the same plasma sample, and a positive correlation between the amount of D antigen on the surface of RBCs and titer was observed after comparison of the titers and MFI(r=0.907;P<0.001): MFI of 38 029±4 756 in 32 titer group, MFI of 83 543±5 537 in

关 键 词：RhD抗原量 Rh分型 效价监测 试剂红细胞 质控 

分 类 号：R457.1[医药卫生—治疗学] R446[医药卫生—临床医学]