中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
Chinese Journal of Medical Genetics
2015年
5期
700-702
,共3页
RHD基因%RHD弱D 101G%RHD弱D type 15%基因测序
RHD基因%RHD弱D 101G%RHD弱D type 15%基因測序
RHD기인%RHD약D 101G%RHD약D type 15%기인측서
RHD gene%RHD weak D 101G%RHD weak D type 15%Sequence analysis
目的 探讨1例罕见的Rh弱D型个体的分子机制.方法 采用常规血清学方法和间接抗球蛋白方法(indirect antiglobulin test,IAT),确认样本RhD血型.应用聚合酶链反应(polymerase chain reaction,PCR)、逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和DNA序列分析等方法对先证者RHD基因转录调控序列和全编码序列进行突变筛选和检测.对cDNA扩增产物进行TA克隆和单倍型分析.结果 先证者RhD血清学检测为弱阳性;PCR扩增实验表明先证者存在RHD基因的第1～10外显子,gDNA和cDNA直接测序101位A/G和845A/G杂合.TA克隆得出两个单倍型,即101A＞G突变(RHD弱D 101G)和845A＞G突变(RHD弱D type 15).结论 第101A＞G突变和845A＞G突变导致先证者红细胞D抗原表达减弱,因而表现为弱D型.
目的 探討1例罕見的Rh弱D型箇體的分子機製.方法 採用常規血清學方法和間接抗毬蛋白方法(indirect antiglobulin test,IAT),確認樣本RhD血型.應用聚閤酶鏈反應(polymerase chain reaction,PCR)、逆轉錄-聚閤酶鏈反應(reverse transcription-polymerase chain reaction,RT-PCR)和DNA序列分析等方法對先證者RHD基因轉錄調控序列和全編碼序列進行突變篩選和檢測.對cDNA擴增產物進行TA剋隆和單倍型分析.結果 先證者RhD血清學檢測為弱暘性;PCR擴增實驗錶明先證者存在RHD基因的第1～10外顯子,gDNA和cDNA直接測序101位A/G和845A/G雜閤.TA剋隆得齣兩箇單倍型,即101A＞G突變(RHD弱D 101G)和845A＞G突變(RHD弱D type 15).結論 第101A＞G突變和845A＞G突變導緻先證者紅細胞D抗原錶達減弱,因而錶現為弱D型.
목적 탐토1례한견적Rh약D형개체적분자궤제.방법 채용상규혈청학방법화간접항구단백방법(indirect antiglobulin test,IAT),학인양본RhD혈형.응용취합매련반응(polymerase chain reaction,PCR)、역전록-취합매련반응(reverse transcription-polymerase chain reaction,RT-PCR)화DNA서렬분석등방법대선증자RHD기인전록조공서렬화전편마서렬진행돌변사선화검측.대cDNA확증산물진행TA극륭화단배형분석.결과 선증자RhD혈청학검측위약양성;PCR확증실험표명선증자존재RHD기인적제1～10외현자,gDNA화cDNA직접측서101위A/G화845A/G잡합.TA극륭득출량개단배형,즉101A＞G돌변(RHD약D 101G)화845A＞G돌변(RHD약D type 15).결론 제101A＞G돌변화845A＞G돌변도치선증자홍세포D항원표체감약,인이표현위약D형.
Objective To explore the molecular basis for an individual with a rare weak D phenotype.Methods Regular serological assaying and indirect antiglobulin testing (IAT) were performed to characterize the RhD blood group.Mutations of the RHD gene were screened by polymerase chain reaction (PCR), reverse transcription PCR and DNA sequencing.Amplified cDNA product was TA cloned and subjected to haplotype analysis.Results The RhD blood group of the proband was determined as weak D.The result of PCR amplification showed that all of the 10 exons of the RHD gene were present.Heterozygote status of 101A/G and 845A/G were determined by gDNA and cDNA sequencing.After TA cloning and haplotype sequencing, two alleles 101A＞G mutation (weak D 101G) and 845G＞A mutation (weak D type 15) were revealed.Conclusion 101A＞G and 845G＞A mutations are responsible for the low expression of RhD antigen on the red blood cells of the proband, which has resulted in a weak D phenotype.
