HLA-DRB1 DNA sequencing based typing: an approach suitable for high throughput typing including unrelated bone marrow registry donors
HLA-DRB1 DNA sequencing based typing: an approach suitable for high throughput typing including unrelated bone marrow registry donors
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DOI：
10.1034/j.1399-0039.2001.057001046.x
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发表时间：
2001-01-01
期刊：
影响因子：
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通讯作者：
Christiansen, F
中科院分区：
文献类型：
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作者：
Sayer, D;Whidborne, R;Christiansen, F
迄今报道的基于人类白细胞抗原-DRB1测序的分型策略需要用一系列组特异性引物对每个样本进行单独扩增，然后对产生的任何聚合酶链式反应(PCR)产物进行测序。虽然这在大多数情况下会导致高分辨率的分型，但也会进行一些不必要的扩增。我在这里报告一种新的方法，即在一个试管中对所有等位基因进行人类白细胞抗原-DRB1第二外显子的扩增。对连续642名西澳大利亚无关骨髓登记供者的回顾分析表明，这种方法在71.1%的病例中得到了明确的分型，如果必要的话，可以通过对原始PCR产物进行一次额外的测序反应来容易地解决多义性。
The HLA-DRB1 sequencing based typing strategies reported to date require separate amplifications of each sample with a series of group-specific primers followed by sequencing of any resulting polymerase chain reaction (PCR) products. Whilst this results in high resolution typing in the majority of cases, a number of unnecessary amplifications are performed Me report here a novel approach where amplification of the second exon of HLA-DRB1 is performed in a single tube for all alleles. Retrospective analysis of 642 consecutive Western Australian unrelated bone marrow registry donors has shown that this approach results in unambiguous typings in 71.1% of cases Ambiguities can be readily resolved if necessary with a single additional sequencing reaction on the original PCR product.