中国人群真实RhD阴性个体和RhDel型RHD基因研究

为观察中国人群真实RhD阴性个体和RhD^el型RHD基因存在情况，采用常规盐水法和抗人球蛋白试验筛选RhD阴性捐血者，通过吸收放散试验性确定其中RhD^el和真实RhD阴性，并采用PCR技术检测RHD基因第4内含子，对吸收放散试验结果与基因检测结果不符的样本进一步采用序列特异性引物PCR(PCR-SSP)技术测定RHD基因第3，4，5，6，7，9和10个外显子，观察基因变异。筛选获得104名RhD阴性捐血者，吸收放散试验鉴定25名（24.0％）为RhD^el型，79名（76.0％）为真实RhD阴性；25例RhD^el全部检测到D基因exon4-intro4-exon5区域，79名真实RhD阴性个体中有5名（6.3％）个体（2名Ccdee，2名ccdEe）检测到D基因第4内含子；PCR-SSP结果显示5名个体中，4名存在全部被检测的D基因区域，1名个体（ccdEe）第5外显子检测阴性。结果表明抗人球蛋白试验确认的RhD阴性中国人存在高比率的RhD^el型，且均可检测出RHD基因第4内含子；若排除RhD^el，携带D基因的RhD阴性中国人的比率明显降低，且这些个体并非如经往报道的均为C抗原阳性。

RHD Gene Investigation of Genuine RhD-negative and RhDel Phenotype Chinese

To investigate RHD gene of RhD el phenotype and genuine RhD negative Chinese, the RhD negative samples were screened by anti globulin test, and the RhD el and genuine RhD negative phenotype were distinguished through absorption/elution method. Subsequently intron 4 of RHD was detected using PCR in all samples. The discrepant samples between absorption/elution test and gene detection were further analyzed by PCR SSP in exon 3, 4, 5, 6, 7, 9 and 10 of RHD gene. As a result, 25 RhD el samples and 79 genuine RhD negative individuals were determined among 104 apparent RhD negative Chinese. All RhD el and 5 (6 3 %) genuine RhD negative individuals (2 CCdee, 2 ccdEe, 1 Ccdee) were detected positive for RHD intron 4. PCR SSP showed that 4 discrepant samples possessed all detected region of RHD gene, and 1 sample (ccdEe) tested negative in only exon 5. It is concluded that the RhD el phenotype is highly detected among apparent RhD negative Chinese, and RHD intron 4 can be detected in all RhD el . The RHD negative Chinese positive for D antigen are lower detected if excluding RhD el phenotype, as well as these individuals are not always having C antigen as previous reports.