中国人RHD基因合子型检测分析  

Determination and analysis of RHD zygosity in Chinese population

作　　者：侯磊[1,2] 郭忠慧[1] 李勤[1] 叶璐夷[1] 朱自严[1] 

机构地区：[1]上海市血液中心,上海200051 [2]华东师范大学生命科学学院

出　　处：《中国输血杂志》2009年第2期110-113,共4页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金(编号:30371312);上海市科技发展基金(编号:074958021);上海市血液中心科研基金(编号:L7-1)共同资助

摘　　要：目的判定中国人群中RHD基因是否是纯合子。方法从上海地区正常献血者中选取RhD阳性、RhD阴性、Del和其它D变异型样本共50份,采用PCR-RFLP方法扩增Rhesus盒子判断RHD合子型,同时相对实时定量real-time quantitative(RQ)PCR扩增检测RHD基因第5、7外显子的特征区域。结果9份样本在2种方法的检测结果中出现矛盾。进一步对RHD基因的特异性序列进行PCR-SSP及克隆测序分析,证实这9份样本中,4份是DⅥⅢ变异型,2份是Del与RHD711del C等位基因杂合子,另外3份是携带有中国RhD阴性人群中较常见的RHD-CE(2—9)-D RH阴性等位基因。结论中国人Rh血型系统遗传背景较白种人复杂,RHD基因存在较多的变异情况。仅使用单一方法判断RHD合子型及RhD血清学表型容易产生误判,而从Rhesus盒子和RHD基因不同外显子2个角度进行综合判断,可以减少误判。Objective To determine whether RHD gene is homozygous in Chinese population. Methods A total of 50 samples, including RhD positive, RhD negative, Del and other D variations from healthy blood donors in Shanghai area were collected. The RHD zygosity was amplified by the PCR-RFLP amplify Rhesus box, and the special sequences of RHD exon 5 and exon 7 were amplified by the relative real-time quantitative PCR （ RQ-PCR ）. Results Discrepancies were found in the 9 samples when the results of the two methods were compared. The further analysis via PCR-SSP and the sequencing of RHD gene showed that, in the 9 discrepant samples, 4 samples were determined to be D^Ⅵ Ⅲ variations, 2 samples were hemizygosity with Del and RHD 711 de/C allele, and 3 samples carried RHD-CE（ 2-9 ）-D RH negative allele, which usually existed in Chinese RhD negative population. Conclusion The molecular bases of Rh blood group system in Chinese population are more complicated than that of the European, so both methods for testing Rhesus box and RHD gene must be used to determine RHD zygosity, in order to avoid inaccurate results.

关 键 词：RHD基因合子型 Rhesus盒子 实时定量PCR 

分 类 号：R457.11[医药卫生—治疗学]