1条突变型RHCE*ce新等位基因的发现  

Identification of a novel and mutant RHCE*ce allele

作　　者：姬艳丽[1] 王贞[1] 魏玲[1] 温机智[1] 赵阳[1] 莫春妍[1] 骆宏[1] 张润青[1] 罗广平[1] 梁华钦[1] 

机构地区：[1]广州血液中心临床输血研究所,广东广州510095

出　　处：《中国输血杂志》2014年第2期155-157,共3页Chinese Journal of Blood Transfusion

基　　金：广东省自然科学基金(S2011040002949);广州省医学科研基金项目(S2011040002949)

摘　　要：目的采用新型Rh血型系统的多重连接依赖式探针扩增(MLPA)检测试剂盒,对广州地区收集的200例RhD阴性献血者进行RHCE基因分型。方法收集RhD阴性献血者200例,从静脉血提取基因组DNA,采用MLPA检测试剂盒进行RHCE基因分型和拷贝数定量分析。对MLPA显示的RHCE拷贝数异常的样本,进行RHCE基因外显子1至10测序分析。对发现的杂合突变进行克隆测序。结果在200例RhD阴性献血者中,MLPA基因分型结果显示有6例标本RHCE等位基因定量拷贝数下降,测序发现RHCE基因外显子2的一个新杂合突变(c.308C>T,p.103Pro>Leu),克隆测序证实了该突变。结论在中国人群中,首次发现了一条新的突变型RHCE*ce(308C>T)等位基因,该等位基因在广州地区RhD阴性献血者中频率为6/400。Objective To analyze the RHCE genotyping in 200 RhD-negative donors from Guangzhou by using the no- vel Multiplex Ligation-dependent Probe Amplification （MLPA） RH genotyping assay kit. Methods Genomic DNAs were extracted from the peripheral blood samples of 200 RhD-negative donors, and the MLPA assay kit was used for the RHCE genotyping and quantitative analysis of the copy numbers of RHCE allele. Then, ten exons of RI-ICE gene were amplified and sequenced in the samples with abnormal copy numbers of RHCE allele in MLPA. Heter0zygous mutations were further ana- lyzed by clone sequencing. Results In the 200 RhD-negative donors, the copy numbers of RHCE allele in 6 samples de- creased in MLPA genotyping assay. A novel heterozygous mutation （ c. 308C 〉 T, p. 103Pro 〉 Leu ） located in exon 2 of RHCE gene was identified, and then this novel mutation was verified by clone sequencing. Conclusion A novel and mu- tant RI＇ICE＊ ce（308C 〉 T） allele was identified （or the first time in the Chinese population,and the frequency of this allele in RhD negative donors from Guangzhou was 6/400.

关 键 词：多重连接依赖的探针扩增(MLPA)技术 RHCE基因 新的突变+型兄眵E^＊ce等位基因 

分 类 号：R457.11[医药卫生—治疗学]