Ucla C, van Rood J J, Gorski J, Mach B
Department of Microbiology, University of Geneva School of Medicine, Switzerland.
J Clin Invest. 1987 Oct;80(4):1155-9. doi: 10.1172/JCI113173.
Recent progress in the molecular genetics of HLA class II antigens has revealed the existence of multiple loci and of a large degree of polymorphism, with more individual alleles than was expected. An accurate detection and analysis of this extensive polymorphism is essential for optimal HLA typing for transplantation and for a reevaluation of HLA-disease association. Because of the limitations of the current typing methods, including restriction fragment length polymorphisms, we have proposed a DNA typing procedure based on hybridization with loci- and allele-specific oligonucleotides. Here we present a much simpler way of analyzing class II micropolymorphism down to the level of single nucleotide differences. RNA oligonucleotide typing (ROT) relies on RNA dot blots and requires 10-20 ml of blood. It is shown that with appropriate oligonucleotide probes, ROT can reliably and unambiguously identify any polymorphism at any of the HLA loci, including new alleles, not identified with previous methods. This illustrates the importance of oligonucleotide typing to optimize HLA matching, in particular for transplantation involving unrelated donors.
人类白细胞抗原（HLA）Ⅱ类抗原分子遗传学的最新进展揭示了多个基因座的存在以及高度的多态性，其个体等位基因数量超出预期。准确检测和分析这种广泛的多态性对于移植的最佳HLA分型以及重新评估HLA与疾病的关联至关重要。由于包括限制性片段长度多态性在内的当前分型方法存在局限性，我们提出了一种基于与基因座和等位基因特异性寡核苷酸杂交的DNA分型程序。在此，我们展示了一种更为简单的分析Ⅱ类微多态性的方法，可精确到单核苷酸差异水平。RNA寡核苷酸分型（ROT）依赖于RNA斑点印迹，仅需10 - 20毫升血液。结果表明，使用合适的寡核苷酸探针，ROT能够可靠且明确地识别任何HLA基因座上的任何多态性，包括先前方法未识别的新等位基因。这说明了寡核苷酸分型对于优化HLA匹配的重要性，特别是对于涉及无关供体的移植。
