流式细胞术评估单采血小板中残余红细胞RhD抗原含量的新方法探讨*

doi:10.3969/j.issn.1671-2587.2026.03.013

临床输血与检验 ›› 2026, Vol. 28 ›› Issue (3): 374-380.DOI: 10.3969/j.issn.1671-2587.2026.03.013

流式细胞术评估单采血小板中残余红细胞RhD抗原含量的新方法探讨^*

张玉宇¹, 范竑佶¹, 魏巧真², 朱心怡¹, 孙娟¹, 向东¹, 孙蕴华¹

¹上海市血液中心,上海 200051;
²安徽医科大学第二附属医院,安徽合肥 230601

收稿日期:2025-09-25 出版日期:2026-06-20 发布日期:2026-07-07
通讯作者: 孙蕴华,主要从事血液保障与用血安全方面研究,（E-mail）sunyunhua@sbc.org.cn。
作者简介:张玉宇,主要从事免疫血液学方面研究,（E-mail）zhangyuyu@sbc.org.cn。
基金资助:
^*本课题受中国输血协会圣湘输血医学发展基金（No.CSBT-SX-2024-06）资助

A Novel Flow Cytometry-based Method for Quantifying Residual RhD Antigen on Red Blood Cells in Apheresis Platelets

ZHANG Yuyu¹, FAN Hongji¹, WEI Qiaozhen², ZHU Xinyi¹, SUN Juan¹, XIANG Dong¹, SUN Yunhua¹

¹Shanghai Blood Center, Shanghai 200051;
²The Second Hospital of Anhui Medical University, Hefei 230601

Received:2025-09-25 Online:2026-06-20 Published:2026-07-07

摘要/Abstract

摘要： 目的建立一种使用流式细胞术检测单采血小板中的残余红细胞RhD抗原量的新方法,评估RhD阴性患者输注RhD阳性血小板的安全性。方法本研究建立一种基于抗体消耗原理的流式细胞术。将倍比稀释的RhD阳性红细胞作为模拟样本,与低效价IgG抗-D抗体反应,通过流式细胞术检测抗体吸收后的剩余量,绘制标准曲线,间接定量红细胞及其碎片的RhD抗原总量。应用该方法检测30例献血者单采血小板样本中残余的红细胞含量。结果单采血小板的红细胞残余量在2×10⁸~1.25×10¹⁰ RBCs/U范围内可有效检出。30例单采血小板样本中,有12例样本的残余红细胞量低于检测范围。18例处于检测范围内的样本中,红细胞残余量大于1×10⁹ RBCs/U的共4例,残余量5×10⁸~1×10⁹ RBCs/U的3例,其余11例为2×10⁸~5×10⁸ RBCs/U。所有样本的红细胞残余量均符合国家标准（低于8×10⁹ RBCs/U）,但显著高于既往报道。结论本研究建立的新方法能够更灵敏、准确地检出血小板制品中残余的红细胞抗原。非Del的RhD阴性患者短期内重复输注RhD阳性血小板制品有产生抗-D抗体风险。

关键词: 流式细胞术, 单采血小板, RhD阴性, 红细胞抗原

Abstract: Objective To establish a novel flow cytometry-based method for quantifying residual RhD antigen on red blood cells (RBCs) in apheresis platelets, aims to assess the immunogenic risk of anti-D immunization in RhD-negative patients who receive RhD-positive platelet transfusion. Methods The method quantified residual RBCs based on anti-D antibody consumption. Simulated samples were prepared by serial dilution of RhD-positive RBCs, then incubated with a low-titer anti-D antibody. After anti-D absorption, the remaining free anti-D was quantitative detected by flow cytometry to generate a standard curve, which correlating antibody test value with RBC count. This method was applied to 30 donor apheresis platelet samples. Results The measurable range of the novel method is 2×10⁸ to 1.25×10¹⁰ RBCs/U. In 12 of the 30 tested samples, the RBC levels were below the limit of detection. In the remaining 18 quantifiable samples, 4 samples contained residual RBC counts>1×10⁹ RBCs/U, 3 samples contained 5×10⁸ to 1×10⁹ RBCs/U, and 11 samples contained 2×10⁸ to 5×10⁸ RBCs/U.All samples met the national standard requirements (<8×10⁹ RBCs/U), while the residual RBC levels were significantly higher than prior reports. Conclusion The novel flow cytometric method offers enhanced sensitivity and accuracy for detecting residual RhD-positive RBCs in platelet products. There is a risk of anti-D alloimmunization in non-DEL RhD-negative patients receiving repeated transfusions of RhD-positive platelet products in a short term.

Key words: Flow cytometry, Apheresis platelet, RhD negative, Erythrocyte antigen

中图分类号:

R457.1

张玉宇, 范竑佶, 魏巧真, 朱心怡, 孙娟, 向东, 孙蕴华. 流式细胞术评估单采血小板中残余红细胞RhD抗原含量的新方法探讨^*[J]. 临床输血与检验, 2026, 28(3): 374-380.

ZHANG Yuyu, FAN Hongji, WEI Qiaozhen, ZHU Xinyi, SUN Juan, XIANG Dong, SUN Yunhua. A Novel Flow Cytometry-based Method for Quantifying Residual RhD Antigen on Red Blood Cells in Apheresis Platelets[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2026, 28(3): 374-380.

参考文献

[1] 国家卫生健康委员会. 全血和成分血使用:WS/T 623—2018[S]. 北京:中国标准出版社,2018.
[2] 英国血液学标准委员会(BCSH),输血特别委员会. 血小板输注指南[J]. 国外医学(输血及血液学分册),2003, 26(5):459-466.
[3] 阳雪新,袁咏梅,吴兆勇,等. ABO次侧不相合血小板输注的有效性和安全性初步研究[J]. 临床输血与检验,2024, 26(1):42-48.
[4] 国家质量监督检验检疫总局,中国国家标准化管理委员会. 全血及成分血质量要求:GB 18469—2012[S]. 北京:中国标准出版社,2012.
[5] Gunson H H,Stratton F,Cooper D G,et al.Primary immunization of Rh-negative volunteers[J]. Br Med J, 1970,1(5696):593-595.
[6] Jakobowicz R,Williams L,Silberman F.Immunization of Rh-negative volunteers by repeated injections of very small amounts of Rh-positive blood[J]. Vox Sang,1972, 23(4):376-381.
[7] 尚红,王毓三,申子瑜,主编. 全国临床检验操作规程[M].4版. 北京:人民卫生出版社,2015:5-7.
[8] Cavagnetto C,Alejo Blanco R,McKenna H,et al. Residual red cells in blood components:A multisite study of fully automated enumeration using a hematology analyzer[J]. Transfusion,2021,61(2):568-578.
[9] Canellini G,Rubin O,Delobel J,et al.Red blood cell microparticles and blood group antigens:an analysis by flow cytometry[J]. Trasfusione Del Sangue,2012, 10(Suppl 2):s39-s45.
[10] 徐文皓,李志强. 血浆置换术治疗高效价抗-D患者效果评价[J]. 中国输血杂志,2011,24(9):752-754.
[11] Thibault L,de Grandmont M J,Cayer M P,et al. Rhesus D antigenic determinants on residual red blood cells in apheresis and buffy coat platelet concentrates[J]. Transfus Med Hemother,2020,47(2):129-134.
[12] Gammon R R,Almozain N,Hermelin D,et al.RhD-alloimmunization in adult and pediatric trauma patients[J]. Transfus Med Rev,2024,38(4):150842.
[13] Cid J,Yazer M H,Lozano M.Platelet transfusion and respecting patient D type[J]. Curr Opin Hematol,2015, 22(6):540-546.
[14] Baldwin M L,Ness P M,Scott D,et al.Alloimmunization to D antigen and HLA in D-negative immunosuppressed oncology patients[J]. Transfusion,1988,28(4):330-333.
[15] Cid J,Carbassé G,Pereira A,et al.Platelet transfusions from D+ donors to D- patients:a 10-year follow-up study of 1014 patients[J]. Transfusion,2011,51(6):1163-1169.
[16] 刘敏,彭涛,于晶晶,等. 肿瘤患者血小板输注致迟发性血清学输血反应的临床特征及输血策略[J]. 中国输血杂志, 2024,37(5):491-494.
[17] Moran C, Zepeda A, Diaz H.Antígeno plaquetario humano:breve revisión del tema y su significado clínico[Human platelet antigen:brief review of the topic and its clinical significance][J]. Rev Med Inst Mex Seguro Soc. 2024,62(suppl 1):1-6.
[18] 郭敏,李建民,刘建辉,等. 去白细胞混合浓缩血小板制备的质量控制及临床应用效果观察[J]. 中国输血杂志, 2023,36(11):1025-1028.
[19] 胡志颖. 单采血小板对血小板质量及输注疗效的临床分析[J]. 现代诊断与治疗,2018,29(24):4018-4020.
[20] Estcourt L J,Birchall J,Allard S,et al.Guidelines for the use of platelet transfusions[J]. Br J Haematol,2017, 176(3):365-394.
[21] 朱雯雯,李莺,宋志群,等. Rh阴性患者输注Rh阳性单采血小板的疗效分析[J]. 临床血液学杂志,2016,29(12):951-953.
[22] 杜霞,李茜,涂源泉,等. 昆明地区RhD初筛阴性献血者RHD基因多态性及分子机制研究[J]. 临床输血与检验, 2026,28(1):97-102.
[23] 张瑛,陈锦荣,林甲进. 22例Del型外显子基因测序分析[J]. 临床血液学杂志,2025,38(10):757-760.
[24] Kitazawa J,Nollet K,Morioka H,et al.Non-D Rh antibodies appearing after apheresis platelet transfusion:stimulation by red cells or microparticles?[J]. Vox Sang, 2011,100(4):395-400.
[25] Coughlan C,Bruce K D,Burgy O,et al.Exosome isolation by ultracentrifugation and precipitation and techniques for downstream analyses[J]. CP Cell Biology,2020,88:e110.
[26] Lai J J,Chau Z L,Chen S Y,et al.Exosome processing and characterization approaches for research and technology development[J]. Adv Sci (Weinh),2022, 9(15):e2103222.
[27] 向东,范亮峰,刘曦,等. 免疫性溶血性输血反应100例分析[J]. 临床输血与检验,2022,24(1):11-15.
[28] Heim M U,Böck M,Kolb H J,et al.Intravenous anti-D gammaglobulin for the prevention of Rhesus isoimmunization caused by platelet transfusions in patients with malignant diseases[J]. Vox Sang,1992, 62(3):165-168.

流式细胞术评估单采血小板中残余红细胞RhD抗原含量的新方法探讨^*

A Novel Flow Cytometry-based Method for Quantifying Residual RhD Antigen on Red Blood Cells in Apheresis Platelets

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