Gao X, Jakobsen I B, Serjeantson S W
John Curtin School of Medical Research, Australian National University, Canberra.
Hum Immunol. 1994 Dec;41(4):267-79. doi: 10.1016/0198-8859(94)90045-0.
This report describes a PCR-based typing protocol for the HLA-A polymorphism. Locus-specific primers selectively amplified HLA-A sequences from exon 1 to exon 3 in a single PCR that avoided co-amplification of other classical and nonclassical class I genes. The allelic variation in exons 2 and 3 of the HLA-A gene was examined with a set of 44 oligonucleotide probes. According to the recognized HLA-A sequences the protocol is potentially able to distinguish all known HLA-A alleles with unique nucleotide sequences in this gene region. The related HLA-A genotypes can also be identified in both homozygous and heterozygous individuals. Thus the protocol provides the highest resolution for HLA-A typing. The PCR-SSO typing technique is accurate, reliable, and particularly suitable for a large number of samples. The DNA typing results from 42 Tenth IHWS B-cell lines are compatible with the serologic and IEF definitions. Sixty-six unrelated donors from a northern Chinese population were also tested, with 16 HLA-A alleles detected. Four subtypes of HLA-A2 were found in this population. The distribution of HLA-A subtypes in the population indicated that 40% of donor-recipient pairs thought to be matched for HLA-A by serology would be mismatched. Two novel HLA-A alleles were identified by unusual oligonucleotide hybridization patterns.
本报告描述了一种基于聚合酶链反应（PCR）的HLA - A多态性分型方案。位点特异性引物在单一PCR反应中从外显子1到外显子3选择性扩增HLA - A序列，避免了其他经典和非经典I类基因的共扩增。使用一组44个寡核苷酸探针检测HLA - A基因外显子2和外显子3中的等位基因变异。根据公认的HLA - A序列，该方案有可能区分该基因区域中所有具有独特核苷酸序列的已知HLA - A等位基因。在纯合子和杂合子个体中也可以鉴定出相关的HLA - A基因型。因此，该方案为HLA - A分型提供了最高分辨率。PCR - SSO分型技术准确、可靠，特别适用于大量样本。42个第十届国际人类白细胞抗原研讨会（IHWS）B细胞系的DNA分型结果与血清学和等电聚焦（IEF）定义相符。还对66名中国北方人群的无关供体进行了检测，检测到16个HLA - A等位基因。在该人群中发现了HLA - A2的4个亚型。该人群中HLA - A亚型的分布表明，40%血清学上认为HLA - A匹配的供体 - 受体对实际上并不匹配。通过异常的寡核苷酸杂交模式鉴定出两个新的HLA - A等位基因。
