山东地区汉族RhD阴性献血者RHD基因的纯合性分析  

Homozygous analysis of RHD gene in RHD negative Han nationality blood donors of Shandong area

作　　者：沈云青[1] 刘小信[2] 张文静[1] 徐群[1] 

机构地区：[1]山东省血液中心,山东济南250014 [2]山东省千佛山医院,山东济南250014

出　　处：《医学检验与临床》2013年第4期3-6,共4页Medical Laboratory Science and Clinics

基　　金：山东省自然科学基金资助

摘　　要：目的：Rhesus血型（简称Rh血型）因其抗原众多、变种各异和临床疾病相关性而备受重视。该血型系统中与临床疾病关联最密切的抗原即RhD抗原具有较高的免疫原性。编码RhD抗原的RHD基因两侧各有一个序列高度相似的Rh盒子基因，RhD阴性即是由上、下游Rh盒子基因之间的基因重组引起。分析RhD阴性孕妇丈夫RHD基因的纯合性可预测胎儿患新生儿溶血病的几率。本研究的目的是分析山东地区汉族人RhD阴性表型形成的分子机制，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阴性献血者的DNA样品首先进行多重聚合酶链反应-序列特异性引物（PCR-SSP）分析。然后对Rh盒子基因进行PCR基因扩增，其扩增产物采用聚合酶链反应-限制性片段长度多态性（PCR-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分析显示存在一个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。结论：HD 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制。RhD阴性个体主要表现为纯合的RHD基因阴性，少部分RhD阴性个体存在杂合的RHD基因和纯合的RHD基因。Objective：The Rh blood group system is one of the most polymorphic and clinical important blood group systems in humans. The highly immunogenic RhD antigen is encoded by RHD gene which is flanked by two Rhesus boxes. While the most common RhD-negativity in Whites is caused by a deletion of RHD between the upstream and downstream Rhesus boxes, the sequence of Rhesus boxes in Chinese is stil unknown. The aim of this study is to analyze the sequence of Rhesus boxes gene and explore the genetic mechanism of the RhD negative phenotype in Chinese Han population. Meanwhile the RHD zygosity can be determined by RFLP analysis of PCR product of Rhesus boxes gene. Methods：DNA of 74 RhD negative samples was firstly analyzed with multiplex PCR based method of specific sequence primer （SSP）. The further analysis is given by Rhesus boxes specific sequencing and the RHD zygosity can be determined by PCR-RFLP analysis of Rhesus boxes gene.Results：46 samples （62%） showed the absence of RHD by PCR-SSP and homozygous RHD negative by PCR-RFLP. 22 samples （30%） showed al RHD specific exons of which 19 were RHD gene hemizygous and three were homozygous for RHD. 5 samples （7%） showed no RHD specific exons but presented one RHD gene by PCR-RFLP analysis and further analysis show at least exons 1 and 10 of RHD with RHD specific PCRs. 1 sample （1%） was positive by multiplex RHD PCR, lacking RHD exon 6. The sequencing of hybrid Rhesus box in 27 RhD negative samples showed normal sequence of hybrid Rhesus box as in Caucasians.Conclusions：The RHD gene deletion is the main mechanism that cause RhD-negativity in Chinese Han population. Chinese RHD gene deletion has taken place within the defined breakpoint region as in Caucasians.

关 键 词：Rhesus血型系统 RHD抗原 RHD基因 基因纯和性 免疫血液学 

分 类 号：R44[医药卫生—诊断学]