【摘要】 目的分析中国汉族人RhD阴性个体Rh盒子基因的序列以阐明中国汉族人群RhD阴性表型形成的分子机理,并对Rh盒子基因的扩增产物进行分析以确定RHD基因的纯合性。方法74例RhD阴性个体的DNA样品首先进行多重聚合酶链反应-序列特异性引物(PCR-sequencespecificprimer,PCR-SSP)分析。然后对Rh盒子基因进行特异性测序分析,同时对Rh盒子基因的扩增产物采用聚合酶链反应-限制性片段长度多态性(restrictfragmentlengthpolymorphism,RFLP)方法进行RHD基因的纯合性测定。结果46例(62%)样品在多重PCR-SSP分析中显示缺失RHD基因,在PCR-RFLP分析中显示为纯合的RHD基因阴性。22例(30%)样品显示存在RHD基因,其中19例显示为杂合的RHD基因,3例显示为纯合的RHD基因。5例(7%)样品缺失RHD基因,但PCR-RFLP分析显示存在1个RHD基因,进一步的分析表明它们至少存在RHD基因第1和10外显子。1例(1%)样品显示存在RHD基因,但缺失第6外显子。对27例在多重PCR分析中显示缺失RHD基因的RhD阴性样品的杂化Rh盒子基因进行DNA测序分析,表明中国人存在与白人相一致的杂化Rh盒子基因序列。结论RHD基因缺失是引起中国汉族人RhD阴性表型形成的主要分子机理。中国人RHD基因缺失发生于与白人相一致的断点区域。更多还原

【Abstract】 Objective To analyze the sequences of Rhesus boxes of RhD gene, and explore the genetic mechanism of RhD negative phenotype in Chinese Han population. Meanwhile the PCR product of Rhesus boxes is analyzed for determining RHD gene homozygosity. Methods DNA of 74 RhD negative samples were firstly analyzed with multiplex PCR-sequence specific primer (SSP). The further analysis was given to Rhesus boxes specific sequencing and RHD gene homozygosity determined by PCR-restriction fragment length polymorphism(RFLP) analysis to Rhesus boxes. Results In DNA samples of 74 RhD negative individuals, 46 samples (62%) showed the absence and homozygous negative of RHD gene; 22 samples (30%) all showed the existence of RHD specific exons, of which 19 were RHD gene heterozygous and 3 were homozygous; regardless of PCR-RFLP analysis showing no RHD specific exons, but further analysis of RHD specific PCR revealed one RHD gene, at least RHD gene exon 1 and 10 existing in 5 DNA samples (7%); 1 sample (1%) was lacking RHD exon 6 although the multiplex PCR showed the RHD gene to be positive. Analyzing the hybrid Rhesus box of 27 RhD negative samples revealed the Han Chinese population to have the same DNA sequence of hybrid Rhesus box as Caucasians. Conclusion The RHD gene deletion is the main molecular mechanism of causing RhD negative formed in Han Chinese population, who have had the RHD gene deletion taken place within the defined breakpoint region as Caucasians.更多还原