高通量MLPA基因分型技术在1个D^(el)表型家系基因鉴定中的应用

High-throughput genotyping MLPA assay in RHD genotyping for a Chinese pedigree with D^(el) phenotype

目的对1个Del表型家系作基因鉴定。方法运用血型血清学方法对先证者及其家系的3名成员作RhD及相关血型抗原鉴定;提取先证者及其父母和妹妹的静脉血DNA。运用高通量红细胞血型基因的多重连接依赖探针扩增技术(MLPA)对先证者及其他成员血型基因作定性及定量分型;用PCR产物直接测序方法对发现的变异作确认。结果血型血清学检测:先证者为Del表型,其他3名家系成员为正常的RhD阳性个体。MLPA基因分型:先证者有2条包含1227G>A突变位点的RHD等位基因,而3名家系成员有1条包含1227G>A突变位点的RHD等位基因及1条正常的RHD等位基因。结论高通量红细胞血型基因分型的MLPA技术能够对Del表型作明确的基因鉴定,有助于节约宝贵的RhD阴性血液。

Objective To determine RHD genotypes for a Chinese pedigree with D^el phenotype. Methods The tradi- tional serological methods including absorption and elution method were used to detect RhD and other blood groups antigens in proband and three fiunily members. Genomic DNAs of the proband and her parents and sister were extracted from periph-eral venous blood. RHD genotypes of the proband and other family members were determined by the novel high-throughput Multiplex Ligation-dependent Probe Amplification （MLPA） as- say qualitatively and quantitatively. Furthermore, the direct se- quencing method was used to determine the identified mutation by MLPA. Results The primary serological tests show the proband with D^el phenotype and the three family members with normal RhD antigen expression. The genotyping results show theproband with two mutant RHD alleles containing 1227G 〉 A mutation and the other family members with one mutant 1227G 〉 A RHD allele and one normal RHD allele. Conclusion High-throughput genotyping MLPA assay can guild the patients with Del phenotype to receive the RhD-positive blood, restricting the use of the rare RhD-negative blood.