A Rapid Method for HLA-typing by Haplotyping

描述（由申请人提供）：用于肾脏和骨髓供体和受体匹配的人类白细胞抗原（HLA）鉴定是移植的强制性要求。许多等位基因组合不能用目前的分型方法来分辨。单倍型特异性提取，HSE，从个体患者的样品中建立单倍型，并允许在不了解家族信息的情况下进行明确的分型。本文的目标是将HSE发展为：1）一种可靠的方法来提高HLA分型。2)为研究人员研究HLA和疾病的影响提供了强大的工具。 在第一阶段，HSE是针对HLA-B开发的。目前有498个HLA-B等位基因与1068个模糊的等位基因组合确定。在9个HLA实验室中成功地证明了HSE用于将HLA-B等位基因分离成单倍体组分。产生了用于分离HLA-B等位基因的常见模糊组合的初步产品，并且HSE在GenoM-6机器人上自动化，用于1至6个单倍体提取的一致的、步行式处理。 根据拟议的第二阶段资金，HSE将扩大用于其他HLA位点，其他HLA连锁基因和主要组织相容性复合体（MHC）中的多态性微卫星标记。第二阶段的另一个目标是证明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.