A Rapid Method for HLA-typing by Haplotyping

描述(由申请人提供)：移植必须鉴定人类白细胞抗原，用于肾脏和骨髓供者和受者的配型。许多等位基因组合不能用目前的分型方法解决。单倍型特异性提取，HSE，从单个患者的样本中建立单倍型，并允许在不知道家族信息的情况下进行明确的分型。这项工作的目标是将HSE发展成为：1)改进人类白细胞抗原配型的可靠方法。2)研究人员研究人类白细胞抗原与疾病关系的有力工具。 在第一阶段，针对人类白细胞抗原-B开发了HSE。目前有1068个含糊的等位基因组合，共鉴定出498个人类白细胞抗原B等位基因。HSE用于将人类白细胞抗原B等位基因分离成单倍体成分已在9个人类白细胞抗原实验室中得到成功验证。产生了用于分离常见的人类白细胞抗原B等位基因歧义组合的初步产品，并在Genom-6机器人上实现了HSE的自动化，以进行1到6个单倍体提取的一致的、可走开的处理。 根据拟议的第二阶段资金，HSE将扩大到用于其他人类白细胞抗原基因座、其他与人类白细胞抗原相关的基因以及主要组织相容性复合体(MHC)中的多态微卫星标记。第二阶段的另一个目标是展示HSE与现有SNP映射信息的直接兼容性，并将HSE的自动化扩展到96井平台。

DESCRIPTION (provided by applicant): Identification of Human Leukocyte Antigens, HLA, for matching of kidney and bone marrow donors and recipients is mandatory for transplantation. Many allele combinations fail to be resolved with current typing methods. Haplotype-Specific Extraction, HSE, establishes haplotypes from an individual patient's sample, and permits the unambiguous typing without knowledge of familial information. It is the goal of the proposed work to develop HSE into: 1) a reliable method to improve HLA typing. 2) a powerful tool for researchers studying the implications of HLA and disease. During Phase I, HSE was developed for HLA-B. There are currently 498 HLA-B alleles identified with 1068 ambiguous allele combinations. The use of HSE for separation of HLA-B alleles into haploid components was successfully demonstrated in nine HLA laboratories. A preliminary product for separation of common ambiguous combinations of HLA-B alleles was produced, and HSE was automated on a GenoM-6 robot, for consistent, walk-away processing for one to six haploid extractions. Under the proposed Phase II funding HSE will be expanded for use at other HLA loci, other HLA-linked genes, and polymorphic micro-satellite markers in the Major Histocompatibility Complex, MHC. It is a further goal of Phase II to demonstrate the direct compatibility of HSE with existing SNP-mapping information and to expand the automation of HSE to a 96-well platform.