Development of high-resolution DNA typing methods for HLA class 1 genes.

1.通过对人类白细胞抗原-A、-B、-C基因座变异的筛选和核苷酸序列分析，发现了一些新的等位基因，包括人类白细胞抗原-B22、-B27、-CW2，以及一些未表达的(空)等位基因。因此，我们可以识别日本人群中存在的大部分(如果不是全部)HLAI类等位基因。采用聚合酶链式反应-聚合酶链式反应方法建立了基于DNA的中等分辨率的人类白细胞抗原-A和-B基因分型试剂盒。此外，还建立了基于DNA的基因分型试剂盒，用于在序列水平上分辨-A2、-A26、-B15和-B40组的人类白细胞抗原-C位点。因此，使用这些聚合酶链式反应-聚合酶链式反应试剂盒可以检测到日语中的大多数人类白细胞抗原I类等位基因。此外，利用聚合酶链式反应-限制性片段长度多态性方法，建立了适用于小样本的、在序列水平上基于DNA的人类白细胞抗原A、B位点分型方法。利用上述方法，我们对日本人群样本中的人类白细胞抗原A、B和C基因座进行了序列水平的分型，并报告了等位基因和单倍型的频率。此外，我们还调查了东亚人群中人类白细胞抗原-B61和-B17组的等位基因和单倍型的多样性。

1. Screening of variations at HLA-A, -B, and -C loci followed by nucleotide sequencing elucidated a number of new alleles including HLA-B22, -B27, -Cw2, and several non-expressed (null) alleles. Consequently, we could identify most, if not all, of the HLA class I alleles existing in the Japanese population.2. DNA-based typing kits for HLA-A and -B loci at the intermediate resolution were established using the PCR-MPH method. Also, DNA-based typing kits for HLA-C locus as well as -A2, -A26, -B15, and -B40 groups at the sequence-level resolution were established using the PCR-MPH method. Thus, most of the HLA class I alleles in Japanese can be detected using these PCR-MPH kits. In addition, DNA-based typing methods for HLA-A and B loci at the sequence-level resolution, suitable for small number of samples, were established using the PCR-RFLP method.3. Using the above methods, we performed sequence-level typing for HLA-A, B, and C loci in Japanese population samples, and reported allele and haplotype frequenecies. Moreover, we investigated the diversity of alleles and haplotypes of HLA-B61 and -B17 groups in East Asian populations.