多重聚合酶链反应分析中国汉族人群RHD基因  

Analysis of RHD gene by multiplex-PCR in Chinese population

作　　者：朱自严[1] 郭忠慧[1] 刘达庄[1] 陆震宇[1] 陈和平[1] 朱俊[1] 张雄民[1] 王健莲[1] Ellenven der Schoot PA Maaskant-van Wijk 

机构地区：[1]上海市血液中心 [2]荷兰红十字血液中心 [3]鹿特丹血液中心

出　　处：《临床输血与检验》2002年第3期5-9,共5页Journal of Clinical Transfusion and Laboratory Medicine

摘　　要：目的　分析中国汉族人群 RHD基因的构成。　方法　采用 2套多重 PCR-SSP扩增体系 ,对 45 0例献血者 (3 2 8例 Rh D阳性、1 1 2例 Rh D阴性、1 0例 D变异型 )的基因组 DNA进行 RHD特异性外显子 3、4、5、6、7、9和内含子 4,RHDΨ及C、c等位基因扩增 ,并与血清学 Rh定型结果进行对比。　结果　 3 2 8例 Rh D阳性献血者中 ,3 2 6例具有全部的 RHD基因特异性外显子 ,另有 2例分别为 DVa和 DIVb;1 1 2例血清学 Rh D阴性献血者中 ,3 5例存在全部的 RHD基因特异性外显子 (3 1 .2 5 % ) ,但该 3 5例样本中未检出 RHDΨ基因 3 7bp的插入片段。 1 0例 D变异型中 3例为 D ,1例 DVa,1例 RHar0 ,1例DDFR;另外 4例中 ,2例缺乏 RHD所有特异性外显子 ,2例扩增出 RHD所有特异性外显子。　结论　中国汉族人群 RHD基因的表达存在多种模式。多重 PCR体系分析 RHD基因既可诊断个体的 Rh D表型 ,又具有确认不完全 D以及发现新的Objective To explore the genomic structure of RHD gene in Chinese Han population \ Methods\ Two sets of multiplex polymerase chain reaction system were used to study genomic DNA from 450 samples of Chinese Han donors (including 328 RhD positive,112 RhD negative and 10 RhD variant identified by serological typing) Exons 3,4,5,6,7,9 and intron 4 of RHD gene were specifically amplified,also C,c alleles of RHCE and the 37 bp insert in RHD Ψ were amplified In addition,Rh system phenotypes of all the samples were examined as a control \ Results\ In RHD DNA types of 328 donors,326 samples were found to be concordant with their D phenotypes which consisted of all RHD specific exons and were typied as RhD positive in serological method,the other two samples showed D IVb and D Va .All RHD specific exons were present in 35 samples (31 25%) out of 112 RhD negative serological typing.None of RHD Ψ was present in these 35 donors with serological RhD typing negative and all RHD specific exons shown by PCR.Among 10 samples of D variant,one D Va ,one R Har 0,one D DFR and three D ⅥⅢ categories were detected.\ Conclusion\ Polymorphisms of RHD gene were present among RhD(-) Chinese Han blood donors.The multiplex PCR assay is reliable in determining genotypes of people who have the D+ and partial D phenotypes as well as in discovering new D variant.

关 键 词：多重聚合酶链反应 中国 汉族人群 RHD基因 RHDΨ基因 不完全D D变异型 

分 类 号：Q754[生物学—分子生物学]