中国人群中发现主要弱D型——弱D15 个体分子背景探究.docVIP

中国人群中发现主要弱D型——弱D15 个体分子背景探究 　 作者：孙国栋， 段现民， 张彦平， 尹志柱， 牛小利， 李艳凤1，赵有良，牛海江 【摘要】 为了探讨中国汉族人群弱D15 型个体的Rh血型系统血型血清学表型及分子背景，采用常规血型血清学技术检测RhD抗原弱阳性个体Rh D、C、c、E、e抗原表型；采用序列特异性引物-聚合酶链反应（SSP-PCR）方法同时检测RHD基因和RHCE基因；标本测序分析RHD基因全长编码区序列；同时通过特异性PCR技术测定RHD合子型或RHD基因数目。结果表明， 血型血清学试验证实为D抗原弱阳性表型的人群中，有18例为弱D15 型（占D抗原弱阳性表型56%），RHD基因全长编码区序列分析发现其第6外显子有一处碱基突变：845Ggt;A，编码区序列其它部分与正常RHD序列相同；检测Rh小因子有3种表型CcEe（2例）、CcEe（ 2例）、ccEe（14例）,分别占弱D15 型的11%、11%、78%，血清学检测结果与分子生物学检测结果一致。RHD杂合性试验鉴定显示仅表型为CcEe的2例标本为纯合型RHD+/RHD+，提示基因型为DCe/DcE；其余为杂合型RHD+/RHD-,提示基因型分别是DCe/dce和DcE/dce。结论： 弱D15型是中国人群中最主要的弱D型，其中大部分为杂合型。 【关键词】 弱D型；弱D15； RHD基因； RHCE基因； 基因突变； 序列分析 　　Molecular Background of Weak D Type 15 as the Predominant Weak D Type Found in Chinese Population 　　AbstractThis study was aimed to investigate the molecular genetic basis and serological phenotype of Rh weak D type 15 individuals. Samples were identified by serological tests and genotyped by sequence specific primer-PCR（SSP-PCR）, and were sequenced to detect the changes of all ten RHD exons. The number of gene RHD was detected through SSP-PCR. The results showed that in tested individuals of weak D type confirmed by the IAT, 18 cases（56% in weak D）were weak D type 15. Rh factors found in 2 weak D type 15 individuals (11%) were C﹢c﹢E﹢e; Rh factors found in 2 weak D type 15 individuals (11%) were C﹢c﹢E-e﹢; others(78%) were c-c﹢E﹢e﹢. The results by serological tests were consistent with the results genotyped by PCR-SSP method. In all 18 samples, the sequencing result revealed a gene mutation 845Ggt;A at the exon 6 of the RHD and the point mutation changed amino acid G282D of the RhD polypeptide. The zygosity test demonstrated that 2 out of 18 weak D type 15 individuals were RHD+/RHD+ homozygous (two DCe/DcE), 16 cases were RHD+/RHD- heterozygous (two DCe/dce and fourteen DcE/dce). It is concluded that Weak D type 15 is predominant in weak D individuals of Chinese Han Nationality, and most of them are heterozygous with various RH haplotypes. 　　Key wordsweak D typ