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Abstract：
OBJECTIVE: HLA-DRB1*16:35 is a low incidence allele in the HLA-DRB1 locus. The objective of this study is to report the ethnicity of DRB1*16:35 and its deduced probable HLA associated haplotype in two Taiwanese unrelated bone marrow hematopoietic stem cell donors and to determine its variation from DRB1*16:02:01 and DRB1*16:01:01.
METHODS: A sequence-based typing method was employed to confirm the low incidence allele DRB1*16:35. Polymerase chain reaction was performed to amplify exon 2 and exon 3 of the HLA-A and HLA-B loci and exon 2 of the HLA-DRB1 locus using group-specific primer sets. The amplicons were sequenced employing BigDye Terminator Cycle Sequencing Ready Reaction kits in both directions according to the manufacturer\'s protocols.
RESULTS: The DNA sequence of DRB1*16:35 is identical to DRB1*16:02:01 in exons 2, except for residue 364 where the C of DRB1*16:02:01 is replaced by the T of DRB1*16:35 (codon 93, CGG->TGG). The nucleotide exchange leads to an amino acid alteration to the protein sequence of DRB1*16:02:01 at residue 93 where the arginine (R) of DRB1*16:02:01 is changed to the tryptophan (W) of DRB1*16:35. We deduced the probable HLA haplotype in association with DRB1*16:35 in Taiwanese to be A*11-B*13- DRB1*16:35.
CONCLUSIONS: Information on the deduced probable HLA haplotype in association with the low incidence DRB1*16:35 allele that we report here is of value for HLA testing laboratories for reference purposes. In addition, it can be used by stem cell transplantation donor search coordinators to determine a strategy for finding compatible donors in unrelated bone marrow donor registries when a patient has this uncommon HLA allele.
METHODS: A sequence-based typing method was employed to confirm the low incidence allele DRB1*16:35. Polymerase chain reaction was performed to amplify exon 2 and exon 3 of the HLA-A and HLA-B loci and exon 2 of the HLA-DRB1 locus using group-specific primer sets. The amplicons were sequenced employing BigDye Terminator Cycle Sequencing Ready Reaction kits in both directions according to the manufacturer\'s protocols.
RESULTS: The DNA sequence of DRB1*16:35 is identical to DRB1*16:02:01 in exons 2, except for residue 364 where the C of DRB1*16:02:01 is replaced by the T of DRB1*16:35 (codon 93, CGG->TGG). The nucleotide exchange leads to an amino acid alteration to the protein sequence of DRB1*16:02:01 at residue 93 where the arginine (R) of DRB1*16:02:01 is changed to the tryptophan (W) of DRB1*16:35. We deduced the probable HLA haplotype in association with DRB1*16:35 in Taiwanese to be A*11-B*13- DRB1*16:35.
CONCLUSIONS: Information on the deduced probable HLA haplotype in association with the low incidence DRB1*16:35 allele that we report here is of value for HLA testing laboratories for reference purposes. In addition, it can be used by stem cell transplantation donor search coordinators to determine a strategy for finding compatible donors in unrelated bone marrow donor registries when a patient has this uncommon HLA allele.
摘要：
目的：HLA-DRB1*16:35是HLA-DRB1位点的低发生率等位基因。这项研究的目的是报告两名台湾无关骨髓造血干细胞供体中DRB1*16：35的种族及其推导的可能的HLA相关单倍型，并确定其与DRB1*16：02：01和DRB1的变异*16：01：01。
方法：采用基于序列的分型方法确认低发生率等位基因DRB1*16:35。使用组特异性引物组进行聚合酶链反应以扩增HLA-A和HLA-B基因座的外显子2和外显子3以及HLA-DRB1基因座的外显子2。根据制造商的方案,采用BigDye终止子循环测序就绪反应试剂盒在两个方向上对扩增子进行测序。
结果:DRB1*16:35的DNA序列与外显子2中的DRB1*16:02:01相同,除了残基364,其中DRB1*16:02:01的C被DRB1*16:35的T取代(密码子93,CGG->TGG)。核苷酸交换导致残基93处DRB1*16:02:01的蛋白质序列的氨基酸改变,其中DRB1*16:02:01的精氨酸(R)变为DRB1*16:35的色氨酸(W)。我们推断台湾与DRB1*16：35相关的可能的HLA单倍型为A*11-B*13-DRB1*16：35。
结论：我们在此报告的与低发病率DRB1*16:35等位基因相关的推断可能HLA单倍型的信息对于HLA检测实验室具有参考价值。此外,干细胞移植供者搜索协调员可以使用该方法来确定当患者具有这种罕见的HLA等位基因时,在无关的骨髓供者登记处寻找相容供者的策略.
方法：采用基于序列的分型方法确认低发生率等位基因DRB1*16:35。使用组特异性引物组进行聚合酶链反应以扩增HLA-A和HLA-B基因座的外显子2和外显子3以及HLA-DRB1基因座的外显子2。根据制造商的方案,采用BigDye终止子循环测序就绪反应试剂盒在两个方向上对扩增子进行测序。
结果:DRB1*16:35的DNA序列与外显子2中的DRB1*16:02:01相同,除了残基364,其中DRB1*16:02:01的C被DRB1*16:35的T取代(密码子93,CGG->TGG)。核苷酸交换导致残基93处DRB1*16:02:01的蛋白质序列的氨基酸改变,其中DRB1*16:02:01的精氨酸(R)变为DRB1*16:35的色氨酸(W)。我们推断台湾与DRB1*16：35相关的可能的HLA单倍型为A*11-B*13-DRB1*16：35。
结论：我们在此报告的与低发病率DRB1*16:35等位基因相关的推断可能HLA单倍型的信息对于HLA检测实验室具有参考价值。此外,干细胞移植供者搜索协调员可以使用该方法来确定当患者具有这种罕见的HLA等位基因时,在无关的骨髓供者登记处寻找相容供者的策略.
