【摘要】 目的 研究分析中山地区初检RhD阴性人群中D变异体血清学和基因分型特征。方法 研究共收集2017年12月～2019年2月中山市人民医院门诊及住院病人的血液样本24 286例，采用微柱凝胶卡法对其中RhD阴性样本进行初筛；再经间接抗人球蛋白试验（indirect antiglobulin test, IAT）确认弱D或部分D变异型；然后用吸收放散试验对剩余样本进行Del表型筛选。同时，通过聚合酶链反应-序列特异性引物（polymerase chain reaction-sequence specific primmer, PCR-SSP）技术对初筛RhD阴性的样本进行RHD等位基因分型以及血清学方法对初筛阴性样本进行C, c, E和e抗原表型的检测。结果 在24 286例血液样本中初筛共检出102例阴性样本，中山地区初检阴性比例约为0.42%。经IAT确认试验共鉴定出7例弱D或部分D血液样本（6.86%），且基因型表现多样，但未见亚洲地区常见的弱D15和弱D12表型。接下来的放散试验检测出25例Del型，通过PCR-SSP基因分型技术又检出3例漏检的Del型，Del型在初检阴性样本中占比27.45%,PCR-SSP结果显示该研究中所有Del型血液样本等位基因均为RHD1227A。该研究纳入的102例初检阴性血液样本的RhCcEe表型分布符合Hardy-Weinberg遗传平衡(χ2=2.625,P>0.05)，表示相应等位基因所占比例在遗传中保持不变。在Del型变异体中，除Ccee和CCee表型外，未见其他表型。结论 中山地区人群RhD变异体有丰富的类型和不同分子机制，其中Del型占比较高，血清学与基因分型结合可提高D变异体的鉴定能力，防止漏检情况的发生。更多还原

【Abstract】 Objective To study serological and genotyping characteristics of RhD gene variants among RhD-negative group in Zhongshan area. Methods Applying miro column gel type card to screen Rh-D-negative samples among 24 286 persontime from December 2017 to February 2019 in Zhongshan People’s Hospital. Then, using indirect anti-globulin method(IAT) to identify partial D phenotype or weak D phenotype, and conducting adsorption and elution test to discern Del phenotype among the above RhD-negative-identified samples. Polymerase chain reaction sequence specific primmer(PCR-SSP) technology was used to analyze RhD-negative samples alleles and RhCcEe phenotype. Results Among the 24 286 blood samples, 102negative samples were initially screened bygelcardsand the proportion of negative samples was about 0.42%. The IAT confirmation test identified 7 cases of weak D or partial D(6.86%), genotypes were diverse, but weak D15 and weak D12commonly seen in Asia were not seen. Then performed absorption and release tests on the remaining negative samples to confirm 25 cases of Del variants. PCR-SSP genotyping technology avoided the missed detection of the other 3 cases of Del variants. The Del type accounted for 27.45%, and the genotyping results showed that all the alleles of the Del type blood samples in this study were RHD1227A. In this study, the RhCcEe phenotype distribution of 102 initial negative samples was consistent with HardyWeinberg genetic balance(χ~2=2.625,P>0.05), indicating that the corresponding allele ratio remained unchanged in inheritance.However, among Del variants, only CCee and Ccee phenotype were observed, and no other phenotypes were found. Conclusion D variants population in Zhongshan area population were various types and in different genetic background. The combination of serology and genotyping can improve the diagnostic ability of RHD variants.更多还原