通过基于序列的高分辨率分型（SBT）揭示的HLA - DQA1和 - DQB1等位基因之间的新型关联。

Novel associations among HLA-DQA1 and -DQB1 alleles, revealed by high-resolution sequence-based typing (SBT).

作者信息

Pera C, Delfino L, Longo A, Pistillo M P, Ferrara G B

机构信息

National Cancer Institute, IST, c/o Advanced Biotechnology Center, Genoa, Italy.

出版信息

Tissue Antigens. 2000 Mar;55(3):275-9. doi: 10.1034/j.1399-0039.2000.550313.x.

DOI:10.1034/j.1399-0039.2000.550313.x

PMID:10777105

Abstract

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: DQA105011 and DQB10201, DQA10505 and DQB103011, DQA101021 and DQB106, DQA101022 and DQB10502.

摘要

尽管基于序列的分型（SBT）是鉴定HLA等位基因的一种有价值的工具，但在用于确定HLA - DQA1和 - DQB1等位基因时存在困难。具体而言，一些HLA - DQA1等位基因在第2外显子的密码子56处有一个三碱基缺失，这会干扰测序读数。此外，常用于HLA - DQB1的引物可能会共扩增HLA - DQB2假基因。为了克服这些问题，我们使用五组特异性聚合酶链反应（PCR）扩增DQA1第2外显子，从而能够分离缺失型和非缺失型DQA1等位基因。使用两组特异性扩增来扩增DQB1第2外显子。为了提高分型分辨率，我们还通过使用序列特异性引物的PCR（PCR - SSP）或SBT分析来分析DQA1的第1、3和4外显子以及DQB1的第3外显子。使用这种方法，我们发现了DQA1和DQB1等位基因之间的一些重要关联：DQA105011和DQB10201、DQA10505和DQB103011、DQA101021和DQB106、DQA101022和DQB10502。