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.
出处
《中国输血杂志》
CAS
CSCD
北大核心
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)
