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首页> 外文期刊>Tissue antigens. >Ambiguities of human leukocyte antigen-B resolved by sequence-based typing of exons 1, 4, and 5.
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Ambiguities of human leukocyte antigen-B resolved by sequence-based typing of exons 1, 4, and 5.

机译：人类白细胞抗原B的歧义可通过基于序列的外显子1、4和5的输入解决。

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摘要

The elucidation of the sequences of human leukocyte antigen-B (HLA-B)-exons 1 through 5 has led to an increase of ambiguities with alleles having identical exon 2 and 3 sequences, but differences in other exons. At the moment, 26 HLA-B alleles show such ambiguities which can be resolved by sequencing the exons in which the differences are located. Here we report a sequence-based typing (SBT) strategy for heterozygous sequencing of exons 1, 4, and 5, in addition to the previously described exons 2 and 3. The strategy was validated against a panel of 25 individuals, carrying HLA-B alleles from 33 different allele groups. Correct assignment of all HLA-B alleles was obtained for exons 1 through 5. In addition, the SBT protocol was used to resolve ambiguities in 50 individuals. The ambiguous combinations studied were B*0705/06, B*0801/19N, B*1512/19, B*180101/17N, B*270502/13/0504, B*350101/42/40N, B*390101/0103, B*400102/0101, B*440201/19N/27, and B*510101/11N/0105/30/32. In all cases, sequencing revealed the first allele to be present, except for three individuals with B*07. One of them typed B*0705; the other two were B*0706. The described SBT protocol for sequencing exons 1, 4, and 5 is a valuable tool for resolving ambiguities of HLA-B alleles with differences in these exons, as well as for studying the polymorphism of HLA-B outside exons 2 and 3.
机译：对人类白细胞抗原-B（HLA-B）-外显子1至5的序列的阐明导致等位基因具有相同的外显子2和3序列，但其他外显子有所不同，这导致歧义性增加。目前，有26个HLA-B等位基因显示出此类歧义，可以通过对差异所在的外显子进行测序来解决。在这里，我们报告了除先前描述的外显子2和3外，对外显子1、4和5进行杂合测序的基于序列的分型（SBT）策略。该策略针对25名携带HLA-B的个体进行了验证来自33个不同等位基因组的等位基因。正确地分配了外显子1至5的所有HLA-B等位基因。此外，使用SBT协议解决了50个个体中的歧义。研究的歧义组合为B * 0705/06，B * 0801 / 19N，B * 1512/19，B * 180101 / 17N，B * 270502/13/0504，B * 350101/42 / 40N，B * 390101/0103 ，B * 400102/0101，B * 440201 / 19N / 27和B * 510101 / 11N / 0105/30/32。在所有情况下，测序均显示出第一个等位基因，但三个B * 07个体除外。其中之一键入B * 0705;其他两个是B * 0706。所描述的用于对外显子1、4和5进行测序的SBT协议是解决这些外显子中具有差异的HLA-B等位基因的歧义以及研究外显子2和3外的HLA-B多态性的宝贵工具。

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