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临床输血与检验

临床输血与检验 ›› 2026, Vol. 28 ›› Issue (1): 97-102.DOI: 10.3969/j.issn.1671-2587.2026.01.015

• 调查研究 • 上一篇    下一篇

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*

杜霞, 李茜, 涂源泉, 陈璐, 张智慧, 彭沫溱, 罗臻, 寸伟, 朱祥明   

  1. 云南昆明血液中心，云南昆明 650506
  • 收稿日期:2025-07-14 发布日期:2026-02-13
  • 通讯作者: 朱祥明，主要从事输血医学研究，（E-mail） kmblood@163.com。
  • 作者简介:杜霞，主要从事免疫血液学研究，（E-mail）2986855626@qq.com。
  • 基金资助:
    *本课题受昆明市卫生科研课题项目（No.2023-11-01-034）资助

Characterization of RHD Gene Polymorphism in Initially Screened D Negative Donors from Kunming Region

DU Xia, LI Qian, TU Yuanquan, CHEN Lu, ZHANG Zhihui, PENG Mozhen, LUO Zhen, CUN Wei, ZHU Xiangming   

  1. Yunnan Kunming Blood Center, Kunming 650506
  • Received:2025-07-14 Published:2026-02-13

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摘要： 目的 研究昆明地区RhD初筛阴性献血者RHD基因的多态性及其分子机制，为建立区域性献血者RHD基因数据库提供数据支持。方法 选择昆明地区2023年11月—2024年8月初筛RhD阴性标本218例，采用间接抗球蛋白试验法（IAT）进行RhD阴性确认，采用盐水试管法进行RhCE表型鉴定。提取全血基因组DNA，采用PCR-SSP法/SSP荧光PCR染料法进行RHD基因分型，对无法确定基因型的标本进行RHD基因1~10外显子Sanger测序分析。结果 检出RhD真阴性表型179例（82.11%），其中RHD*01N.01（RHD全缺失）型154例（86.03%），表型以ccee为主（87.01%）;携带非功能性RHD等位基因25例（13.97%），包括RHD*01N.03 20例、RHD*01N.16 3例、RHD*01N.05 1例、RHD*01N.59 1例，表型以Ccee为主（64%）。检出D变异型39例（15.89%），其中RHD*DEL1(c.1227G>A)型34例，表型均为C抗原阳性（Ccee 27例，CCee 7例）;弱D/部分D型4例，包括RHD*DVI.3 2例、RHD*weak D type 71 1例、RHD*weak D type108 1例;另检出1例RHD*01/RHD*01N.01，基因型与血清学表型结果不一致。RHD*01N.01女性献血者不规则抗体（主要为抗-D）阳性率9.84%。结论 昆明地区RhD初筛阴性献血者RHD基因多态性显著，RhD真阴性比例高于国内部分地区，D变异型以“亚洲型”DEL为主，比例低于国内部分地区。研究结果为本地区RhD阴性和D变异型个体精准输血提供了理论和数据支持。

关键词: RhD阴性, D变异型, 亚洲型DEL, 基因多态性, 分子机制

Abstract: Objective To investigate the polymorphism of the RhD gene and its molecular mechanism in RhD-negative blood donors in Kunming region, and to provide data support for establishing a regional RHD gene database for blood donors. Methods A total of 218 RHD-negative samples were enrolled from November 2023 to August 2024. RhD negativity was confirmed by the Indirect Antiglobulin Test (IAT), and the RhCE phenotypes were identified using the saline test tube method. Genomic DNA was extracted from whole blood samples. RHD genotyping was performed via PCR-SSP method /SSP fluorescent PCR dye methods. For samples with undetermined genotypes, Sanger sequencing analysis of exons 1~10 of the RHD gene was conducted. Results A total of 179 cases (82.11%) were identified as serologically confirmed RhD-negative phenotypes, among which 154 cases (86.03%) carried the RHD*01N.01 genotype (complete RHD deletion) , with the predominant phenotype being ccee (87.01%). Twenty-five cases (13.97%) harbored non-functional RHD alleles, including RHD*01N.03 (20 cases), RHD*01N.16 (3 cases), RHD*01N.05 (1 cases), and RHD*01N.59 (1 cases). The main phenotype was Ccee (64%). Thirty-nine cases (15.89%) were classified as D variants, among which 34 cases belonged to the RHD*DEL1(c.1227G>A) type, all of which expressed the c antigen (comprising 27 Ccee and 7 CCee phenotypes). Four cases were identified as weak D/partial D types, including RHD* VI.3 (2 cases), RHD*weak D Type 71 (1 cases), and RHD*weak D Type108 (1 cases). Additionally, 1 case with the RHD*01/RHD*01N.01 genotype was detected, showing inconsistency between genotype and serological phenotype. The seropositivity rate of irregular antibodies (predominantly anti-D) in female blood donors with the RHD*01N.01 genotype was 9.84%. Conclusion The RhD gene exhibits significant polymorphism among initially screened RhD-negative donors in Kunming region. The proportion of true RhD-negative donors is higher than that in some other regions. The Asian-type DEL is the predominant D variant, though its overall proportion is relatively low. This study provides theoretical and data support for precision blood transfusion in individuals with RhD-negative and D variants.

Key words: RhD negative, D variants, Asian-type DEL, Gene polymorphism, Molecular mechanism

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