Abstract：
OBJECTIVE: HLA-C*04:82 is a low incidence HLA-C allele. The aim here is to report the ethnicity of C*04:82 and its associated HLA haplotypes among Taiwanese individuals.
METHODS: A sequence-based typing method was used to confirm this low incidence allele. Polymerase chain reaction was performed to amplify exons 2 and 3 of the HLA-A, HLA-B, and HLA-C loci and exon 2 of the HLA-DRB1 locus using group-specific primer sets. The amplicons were sequenced in both directions using Terminator Cycle Sequencing Ready Reaction kits and the manufacturer\'s protocols. The potential unrelated bone marrow stem cell donors in this study were randomized individuals with Taiwanese ethnicity who participated in the Tzu Chi Bone Marrow Donor Registry. The family members in the family part of the study were volunteer blood donors.
RESULTS: The DNA sequence of C*04:82 was identical to C*04:01:01:01 in exons 2, 3, and 4. It differed from C*04:01:01:01 in exon 5 where a segment of nucleotides (CTAGCTGTC) was inserted between residues 969 and 970 of C*04:01:01:01. The insertion of these nucleotides caused a 35 amino acid alteration to the protein sequence of C*04:01:01:01. Three probable HLA haplotypes that were associated with C*04:82 among Taiwanese individuals were deduced. Confirmation of the DNA and protein sequences of C*04:82 and its Taiwanese ethnicity were established in this study.
CONCLUSIONS: The ethnicity of the C*04:82 allele and the deduced probable HLA haplotypes associated with the low-incidence C*04:82 allele are of value for reference purposes for HLA testing laboratories. In addition, they can be used by search coordinators to aid the creation of a strategy for finding compatible stem cell donors for patients who carry this uncommon HLA allele.
METHODS: A sequence-based typing method was used to confirm this low incidence allele. Polymerase chain reaction was performed to amplify exons 2 and 3 of the HLA-A, HLA-B, and HLA-C loci and exon 2 of the HLA-DRB1 locus using group-specific primer sets. The amplicons were sequenced in both directions using Terminator Cycle Sequencing Ready Reaction kits and the manufacturer\'s protocols. The potential unrelated bone marrow stem cell donors in this study were randomized individuals with Taiwanese ethnicity who participated in the Tzu Chi Bone Marrow Donor Registry. The family members in the family part of the study were volunteer blood donors.
RESULTS: The DNA sequence of C*04:82 was identical to C*04:01:01:01 in exons 2, 3, and 4. It differed from C*04:01:01:01 in exon 5 where a segment of nucleotides (CTAGCTGTC) was inserted between residues 969 and 970 of C*04:01:01:01. The insertion of these nucleotides caused a 35 amino acid alteration to the protein sequence of C*04:01:01:01. Three probable HLA haplotypes that were associated with C*04:82 among Taiwanese individuals were deduced. Confirmation of the DNA and protein sequences of C*04:82 and its Taiwanese ethnicity were established in this study.
CONCLUSIONS: The ethnicity of the C*04:82 allele and the deduced probable HLA haplotypes associated with the low-incidence C*04:82 allele are of value for reference purposes for HLA testing laboratories. In addition, they can be used by search coordinators to aid the creation of a strategy for finding compatible stem cell donors for patients who carry this uncommon HLA allele.
摘要：
目的:HLA-C*04:82是低发生率的HLA-C等位基因。这里的目的是报告台湾个体中C*04:82的种族及其相关的HLA单倍型。
方法：使用基于序列的分型方法来确认该低发生率等位基因。进行聚合酶链反应以扩增HLA-A的外显子2和3，HLA-B,以及使用组特异性引物组的HLA-C基因座和HLA-DRB1基因座的外显子2。使用终止子循环测序就绪反应试剂盒和制造商的方案在两个方向上对扩增子进行测序。本研究中潜在的无关骨髓干细胞捐献者是参与慈济骨髓捐献者注册的台湾种族的随机个体。研究的家庭部分中的家庭成员是自愿献血者。
结果:C*04:82的DNA序列在外显子2、3和4中与C*04:01:01相同。它不同于外显子5中的C*04:01:01:01,其中核苷酸区段(CTAGCTGTC)插入在C*04:01:01:01的残基969和970之间。这些核苷酸的插入导致C*04:01:01:01的蛋白质序列发生35个氨基酸的改变。推断了台湾个体中与C*04:82相关的三种可能的HLA单倍型。在这项研究中，确定了C*04：82的DNA和蛋白质序列及其台湾种族。
结论：C*04:82等位基因的种族和与低发生率C*04:82等位基因相关的推测可能的HLA单倍型对于HLA检测实验室具有参考价值。此外,搜索协调员可以使用它们来帮助创建一种策略,为携带这种罕见HLA等位基因的患者寻找相容的干细胞供体.
方法：使用基于序列的分型方法来确认该低发生率等位基因。进行聚合酶链反应以扩增HLA-A的外显子2和3，HLA-B,以及使用组特异性引物组的HLA-C基因座和HLA-DRB1基因座的外显子2。使用终止子循环测序就绪反应试剂盒和制造商的方案在两个方向上对扩增子进行测序。本研究中潜在的无关骨髓干细胞捐献者是参与慈济骨髓捐献者注册的台湾种族的随机个体。研究的家庭部分中的家庭成员是自愿献血者。
结果:C*04:82的DNA序列在外显子2、3和4中与C*04:01:01相同。它不同于外显子5中的C*04:01:01:01,其中核苷酸区段(CTAGCTGTC)插入在C*04:01:01:01的残基969和970之间。这些核苷酸的插入导致C*04:01:01:01的蛋白质序列发生35个氨基酸的改变。推断了台湾个体中与C*04:82相关的三种可能的HLA单倍型。在这项研究中，确定了C*04：82的DNA和蛋白质序列及其台湾种族。
结论：C*04:82等位基因的种族和与低发生率C*04:82等位基因相关的推测可能的HLA单倍型对于HLA检测实验室具有参考价值。此外,搜索协调员可以使用它们来帮助创建一种策略,为携带这种罕见HLA等位基因的患者寻找相容的干细胞供体.
