Lazaro Ana, Tu Bin, Yang Ruyan, Xiao Yi, Kariyawasam Kanthi, Ng Jennifer, Hurley Carolyn Katovich
CW Bill Young Marrow Donor Recruitment and Research Program, Department of Pediatrics, Georgetown University Medical Center, Rockville, MD, USA.
Methods Mol Biol. 2013;1034:161-95. doi: 10.1007/978-1-62703-493-7_9.
DNA sequencing is a powerful technique for identifying allelic variation within the human leukocyte antigen (HLA) genes. Sequencing is usually focused on the most polymorphic exons of the class I (HLA-A, -B, -C) and class II (HLA-DR, -DQ, and -DP) genes. These exons encode the antigen recognition site, the region of the HLA molecule that binds peptides and interacts with the T cell receptor for antigen and natural killer cell immunoglobulin-like receptors (KIR). Sanger sequencing of amplified DNA from each HLA gene from a preparation containing one or two alleles yields a sequence that is used to identify the alleles by comparison with a reference database.
DNA测序是一种用于识别人类白细胞抗原(HLA)基因内等位基因变异的强大技术。测序通常聚焦于I类(HLA-A、-B、-C)和II类(HLA-DR、-DQ和-DP)基因中多态性最高的外显子。这些外显子编码抗原识别位点,即HLA分子中结合肽段并与抗原T细胞受体及自然杀伤细胞免疫球蛋白样受体(KIR)相互作用的区域。对含有一个或两个等位基因的样本中每个HLA基因的扩增DNA进行桑格测序,可得到一个序列,通过与参考数据库比较来识别等位基因。