Novel associations among HLA-DQA1 and -DQB1 alleles, revealed by high-resolution sequence-based typing (SBT).
Novel associations among HLA-DQA1 and -DQB1 alleles, revealed by high-resolution sequence-based typing (SBT).
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高分辨率序列分型 (SBT) 揭示了 HLA-DQA1 和 -DQB1 等位基因之间的新关联。
DOI：
10.1034/j.1399-0039.2000.550313.x
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发表时间：
2000
期刊：
影响因子：
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通讯作者：
G. Ferrara
中科院分区：
文献类型：
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作者：
C. Pera;L. Delfino;A. Longo;M. Pistillo;G. Ferrara
尽管基于序列的分型（SBT）是一种有价值的HLA等位基因鉴定工具，但在用于确定HLA-DQA 1和-DQB 1等位基因时存在困难。具体地，一些HLA-DQA 1等位基因在外显子2的密码子56处具有三个碱基缺失，这干扰测序读数。此外，HLA-DQB 1常用的引物可能会共同扩增HLA-DQB 2假基因。为了克服这些问题，我们扩增DQA 1外显子2使用5组特异性聚合酶链反应（PCR），允许从非删除DQA 1等位基因删除分离。使用两个组特异性扩增来扩增DQB 1外显子2。为了提高分型分辨率，我们还分析了DQA 1外显子1，3和4和DQB 1外显子3通过PCR使用序列特异性引物（PCR-SSP）或SBT分析。使用该方法，我们发现了DQA 1和DQB 1等位基因之间的一些重要关联：DQA 1 *05011和DQB 1 *0201，DQA 1 *0505和DQB 1 *03011，DQA 1 *01021和DQB 1 *06，DQA 1 *01022和DQB 1 *0502。
Althought it is a valuable tool for the identification of HLA alleles, sequence-based typing (SBT) presents difficulties when used to determine HLA-DQA1 and -DQB1 alleles. Specifically, some HLA-DQA1 alleles have a three-base deletion at codon 56 of exon 2 that interferes with the sequencing read. Moreover, the frequently used primers for HLA-DQB1 may co-amplify the HLA-DQB2 pseudogene. To overcome these problems, we amplified DQA1 exon 2 using five group-specific polymerase chain reactions (PCRs) which allowed separation of deleted from non-deleted DQA1 alleles. DQB1 exon 2 was amplified using two group-specific amplifications. To increase typing resolution, we also analyzed DQA1 exons 1, 3 and 4 and DQB1 exon 3 by PCR using sequence-specific primers (PCR-SSP) or SBT analysis. Using this method we found some important associations between DQA1 and DQB1 alleles: DQA1*05011 and DQB1*0201, DQA1*0505 and DQB1*03011, DQA1*01021 and DQB1*06, DQA1*01022 and DQB1*0502.