DNA测序分析鉴定2个弱D新等位基因

Identification of two novel weak D alleles by sequence-based typing

目的研究D变异的分子基础,探讨弱D新等位基因产生的分子遗传机制。方法采用血清学方法筛查D变异体,D变异体采用聚合酶连反应进行RHD基因的全编码区进行扩增并进行直接测序。杂交合子技术检测RHD的杂合性。结果血清学检测为弱D表型,DNA序列分析检测到2个新等位基因,1个是weak D 399C新等位基因,第3外显子有1个c.399G>C突变,导致编码133位氨基酸由赖氨酸变成天冬酰胺(p.K133N)。另1个是weak D 1102A新等位基因,在2个标本中检测到第8外显子c.1102G>A突变,导致编码368位氨基酸由甘氨酸变成精氨酸(p.G368R)。结论本研究采用基因测序技术检测和鉴定RHD变异体,并鉴定2个新等位基因。

Objective To study the molecular basis of D variant and explore the molecular genetic mechanism of novel weak D alleles.Methods Blood samples were screened for D variants by serological method.The nucleotide sequences of coding region were amplified by PCR and sequenced directly,and RHD gene heterozygosity was detected.Results Weak D phenotype was confirmed by serological test,and two novel alleles were detected by DNA sequencing.The first was novel weak D 1102A allele,1102G>A mutation in exon 8,resulting in a 368Glu>Arg substitution in two samples.The second was novel weak D 399C allele,carried a 399G>C mutation in exon 3,which led to a 133Lys>Asn substitution.Conclusion In this study,D variants were detected by sequence-based typing,and two new alleles were identified.