Molecular basis for D- Japanese: identification of novel DEL and D- alleles
Molecular basis for D- Japanese: identification of novel DEL and D- alleles
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DOI：
10.1111/vox.12290
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发表时间：
2015-11-01
期刊：
影响因子：
2.7
通讯作者：
Tadokoro, K.
中科院分区：
文献类型：
--
作者：
Ogasawara, K.;Suzuki, Y.;Tadokoro, K.
背景和目的日本人D-的发生率约为0-50/0，但与高加索人群相比，明显的D-个体的DEL相对较常见。方法采用标准的血清学方法，对3526例D血型献血者进行RhD分型。从全血中提取基因组DNA，进行聚合酶链式反应(PCR)和测序。多重聚合酶链式反应检测所有RIID外显子，并使用[‘CR-序列特异性引物(PCR-SSP)]检测RIID缺失(Ri/D*0IN.01)和C.1227G>；结果3526例D型个体中，3091例(87-7%)为RHD*011V.01纯合子，318例(9-00)例为RHD*DIEL.01/RHD*01 N.01或RHD*0 IEL.01/RHD*01 EL.01。在3526个个体中，有103个个体(2-90/n)存在D-CE-D杂合等位基因RIID*OIN.04，观察到RIICE*Ce与RIID*01EL.01和RIID*01N.04的关联。其余14个个体携带1？111)*0IN.01杂合子，具有以下等位基因之一：RIID*01N.06(3)，RIID*01N.07(1)，RIID*04N.01(1)，RIILYDEL8(1)，RIID与c.761C>；G(p.Ser254Ter)(2)，RIID与c.1252T>；A(p.Ter418Lyscx26)(2)和明显共同的RIID(4)。抗-D吸附和洗脱试验显示C.761C和GT；G突变的个体为D，而C.1252T和GT；A突变的个体为DEL。结论用常规的聚合酶链式反应-单链构象多态性分析可以检测到96%以上的日本D的RI-ID基因。此外，我们还发现了一个新的具有C.1252T和gt；A突变的Del等位基因和一个新的具有c.761C>；G无义突变的RI-ID沉默等位基因。
Background and Objectives The occurrence of D- is approximately 0-50/0 in Japanese, but DEL in apparently D- individuals is relatively common compared with that in Caucasian populations. On the basis of molecular genetics, we examined D- Japanese blood donors.Methods A standard serological technique was used for RhD typing, and we selected 3526 D- blood samples. Genomic DNA obtained from whole blood was used for RIID analysis by polymerase chain reaction (PCR) and sequencing. Multiplex PCR to detect all of the RIID exons and use of ['CR-sequence-specific primer (PCR-SSP) to detect RIID deletion (Ri/D*0IN.01) and c.1227G>A mutation (for RHD*0 I ELM]) were performed.Results Multiplex PCR and PCR-SSP revealed that 3091 of 3526 D individuals (87-7%) were homozygous for RHD*011V.01, and 318 individuals (9-00,/n) had the RHD*DIEL.01/RHD*0 1 N.01 or RHD*0 IEL.0 1/RHD*0 1 EL.01 genotype. The other 103 in the 3526 individuals (2-90/n) had the known D-CE-D hybrid allele, RIID*OIN.04, and the association of RIICE*Ce with RIID*01EL.01 as well as RIID*01N.04 was observed. The remaining 14 individuals had 1?111)*0 IN.01 hernizygous with one of the following alleles: RIID*01N.06 (3), RIID*01N.07 (1), RIID*04N.01 (1), RIILYDEL8 (1), RIID with c.761C>G (p.Ser254Ter) (2), RIID with c.1252T>A (p.Ter418Lyscx26) (2) and apparently common RIID (4). Adsorption and elution tests with anti-D revealed that the individuals with c.761C>G mutation were D while the individuals with c.1252T>A mutation were DEL.Conclusions The RI-ID genotype of more than 96% of D Japanese could be determined by conventional PCR-SSP. In addition, we identified a novel DEL allele having c.1252T>A mutation and a novel RI-ID silencing allele having c.761C>G nonsense mutation.