中国人群RhD阴性个体中D基因多态性的研究

目的 建立RHD基因分型技术，分析中国人群RhD阴性个体的RHD基因多态性分布状况。方法 设计8对特异性引物扩增RHD7个外显子(3，4，5，6，7，9，10)和内含子4，应用PCR—SSP技术，对l15例经常规血清学试验和吸收放散试验鉴定的“Rh阴性个体”，进行D基因多态性的研究。结果 l15例RhD阴性个体，用吸收放散试验检测后分成2组，一组被确认为RhD阴性表现型共88例(76．5％)，另一组是Del(放散)表现型共27例(23．5％)。应用PCR—SSP法对l15例RhD阴性个体作RHD内含于4检查，结果Del型个体都显示有RHD内含于4；对l15例中的17例进一步作RHD的7个外显子的鉴定，其中6例Del带有完整的RHD基因，ll例经吸收放散试验确认为RhD阴性表现型的个体则显示4种情况：7例全部缺失RHD基因，2例(1例ccEe和1例Ccee)带有完整的RHD基巴，1例缺失RHD的第5外显子和1例仅携带第10外显子。结论 PCR—SSP技术可用于RHD基因的研究。常规血清学鉴定的RhD阴性中存在较高比例的Del型，且有可能带有完整的RHD基因。而被吸收放散试验确认为RhD的阴性个体有可能携带RHD基因并显示出多态性，这些个体中有Rhc抗原表达，有别于献中认为全局RhC抗原表达的报告。

Study on RHD gene polymorphism among the apparently RhD-negative Chinese

Objectives To establish the method of RHD genotyping by PCR SSP and preliminary study on RHD gene polymorphism among apparently RhD negative Chinese.Methods The polymerase chain reaction with sequence specific primers (PCR SSP) based RhD genotyping method has been developed using eight RHD specific primers designed to amplify RHD exon 3, 4, 5, 6, 7, 9, 10 and intron 4.Using the PCR SSP method,the RHD gene polymorphism was studied among 115 cases of apparently RhD negative Chinese who were identified by the routine serological methods and absorption elution tests.Results Among 115 RhD negative Chinese,88 samples (76.5%) were confirmed as RhD negative and 27 samples (23.5%) were Del phenotype by the absorption elution tests.Using PCR SSP method,RHD intron 4 were detected in all Del individuals.17 individuals from the 115 cases were studied for seven exons of RHD,6 Del cases were found to carry intact RHD gene.11 cases that confirmed as RhD negative showed 7 cases with total deletion of RHD gene,2 cases with intact RHD gene,1 case with the deletion of RHD exon 5 and 1 case carrying only RHD exon 10.Conclusions The PCR SSP could be used to study RHD genotype.Del phenotypes were highly detected among the apparently RhD negative Chinese individuals and Del cases had the possibility of carrying the intact RHD gene.The apparently RhD negative Chinese individuals identified also had the possibility of carrying the RHD gene and showed the RHD gene polymorphism.The RhD negative Chinese carried RHD gene were found the RHc phenotype which was different from the reported all RhC phenotypes among the RhD negative Chinese.